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  • Electronic Resource  (28)
  • 1990-1994  (24)
  • 1955-1959  (4)
  • 1920-1924
  • 1890-1899
  • gene transfer
  • 1
    Electronic Resource
    Electronic Resource
    Transformation of wheat (Triticum aestivum L.) through electroporation of protoplasts (1994)
    Springer
    Plant cell reports 14 (1994), S. 192-196 
    Details
    ISSN: 1432-203X
    Keywords: gene transfer ; selection for phosphinothricin resistance ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts isolated from embryogenic suspension cultures of wheat (Triticum aestivum cv. Hartog) were electroporated in the presence of plasmid pEmuGN and/or pEmuPAT, which contained the reporter gene gus and selectable marker gene bar, respectively. Under optimised electroporation conditions, up to 0.9% of viable protoplasts displayed gus activity two days after electroporation. To select for phosphinothricin (PPT) resistant colonies, electroporated protoplasts were incubated for six weeks in a medium containing 10 μg/ml PPT. The cells surviving the selection were maintained as individual colonies on solid medium or as suspension cultures. More than 60% of these colonies exhibited tolerance to 40 μg/ml PPT when tested 10 months after initial selection. To date, 57 green plants have been regenerated from these colonies and 24 have been transferred to soil. Southern blot analyses of colonies and plants, using the bar gene sequence as the probe, confirmed transformation of the cells. Positive PAT assays of both regenerated colonies and plants indicated the presence of the bar gene product. These results provide a basis for the establishment of routine procedures for transformation of wheat by direct gene transfer into protoplasts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233789
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  • 2
    Electronic Resource
    Electronic Resource
    Agrobacterium-mediated transformation of commercial melon (Cucumis melo L., cv. Amarillo Oro) (1994)
    Springer
    Plant cell reports 13 (1994), S. 145-148 
    Details
    ISSN: 1432-203X
    Keywords: muskmelon ; gene transfer ; Agrobacterium ; regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cotyledon explants of muskmelon (Cucumis melo L., cv. Amarillo Oro) seedlings were co-cultivated with disarmed Agrobacterium tumefaciens strain LBA4404 that contained the binary vector plasmid pBI121.1. The T-DNA region of this binary vector contains the Nopaline synthase/neomycin phosphotransferase II (NPTII) chimeric gene for kanamycin resistance and the Cauliflower Mosaic Virus 35S/β-glucuronidase (GUS) chimeric gene. After infection, the cotyledon pieces were placed in induction medium containing 100 mg/l kanamycin. Putative transformed shoots were obtained, followed by the development of morphologically normal plantlets. The transgenic nature of regenerants was demonstrated by polymerase chain reaction, Southern blot analysis, plant growth on medium selective for the transgene (NPTII) and expression of the co-transformed GUS gene. Factors affecting the transformation procedure are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00239881
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  • 3
    Electronic Resource
    Electronic Resource
    Fertile transgenic barley by particle bombardment of immature embryos (1994)
    Springer
    Plant molecular biology 24 (1994), S. 317-325 
    Details
    ISSN: 1573-5028
    Keywords: fertile transgenic barley ; gene transfer ; Hordeum vulgare ; neomycin phosphotransferase II ; particle bombardment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic, fertile barley (Hordeum vulgare L.) from the Finnish elite cultivar Kymppi was obtained by particle bombardment of immature embryos. Immature embryos were bombarded to the embryonic axis side and grown to plants without selection. Neomycin phosphotransferase II (NPTII) activity was screened in small plantlets. One out of a total of 227 plants expressed the transferred nptII gene. This plant has until now produced 98 fertile spikes (T0), and four of the 90 T0 spikes analyzed to date contained the nptII gene. These shoots were further analyzed and they expressed the transferred gene. From green grains, embryos were isolated and grown to plantlets (T1). The four transgenic shoots of Toivo (the T0 plant) produced 25 plantlets as T1 progeny. Altogether fifteen of these T1 plants carried the transferred nptII gene as detected with the PCR technique, fourteen of which expressed the nptII gene. The integration and inheritance of the transferred nptII gene was confirmed by Southern blot hybridization. Although present as several copies, the transferred gene was inherited as a single Mendelian locus into the T2 progeny.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00020170
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  • 4
    Electronic Resource
    Electronic Resource
    Expression of a humanS-adenosylmethionine decarboxylase cDNA in transgenic tobacco and its effects on polyamine biosynthesis (1994)
    Springer
    Transgenic research 3 (1994), S. 26-35 
    Details
    ISSN: 1573-9368
    Keywords: gene transfer ; polyamines ; S-adenosylmethionine decarboxylase ; tobacco ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract S-adenosylmethionine decarboxylase (SAMDC; EC 4.1.1.50) is a key regulatory enzyme in the polyamine biosynthetic pathway. Numerous studies have shown that the enzyme activity and polyamine levels are generally correlated with cellular growth in plants, animals and bacteria. In order to gain more insight into the role of polyamines in plants, human SAMDC cDNA under control of the 35S promoter of cauliflower mosaic virus, along with a neomycin phosphotransferase gene, was transferred to tobacco (Nicotiana tabacum cv. Xanthi) viaAgrobacterium tumefaciens. Transgenic plants showed the presence of human SAMDC mRNA and a 2-4-fold increase in SAMDC activity. In the transformed tissues, putrescine levels were significantly reduced, while spermidine content was 2–3 times higher than the control tissues. Cellular spermine content was either increased or remained unchanged. Excised leaf segments from transformed plants frequently produced shoots even on callus inducing medium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01976024
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  • 5
    Electronic Resource
    Electronic Resource
    Metabolic engineering of plant secondary products (1994)
    Springer
    Transgenic research 3 (1994), S. 109-115 
    Details
    ISSN: 1573-9368
    Keywords: gene transfer ; metabolic engineering ; overexpression ; secondary metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plants interact with their environment by producing a diverse array of secondary metabolites. Many of these compounds are valued for their medicinal, industrial or agricultural properties. Other secondary products are toxic or otherwise undesirable and can reduce the commercial value of crops. Gene transfer technology offers new opportunities to modify directly plant secondary product synthesis through metabolic engineering. This article reviews some of the strategies which have been used to increase or decrease the synthesis of specific plant metabolites, as well as methods for expanding the biosynthetic capabilities of individual species.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01974088
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  • 6
    Electronic Resource
    Electronic Resource
    An assessment of gene transfer by pollen from field-grown transgenic potatoes to non-transgenic potatoes and related species (1994)
    Springer
    Transgenic research 3 (1994), S. 216-225 
    Details
    ISSN: 1573-9368
    Keywords: Solanum tuberosum ; genetic modification ; transformation ; gene transfer ; genetic isolation ; risk assessment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Information on the extent of transgene dispersal by pollen to adjacent potato plots and to related weed species is an important requisite for risk assessment; a procedure followed before novel transgenic plants are evaluated under field conditions. The purpose of the investigation was to determine the frequency of cross-pollination between potato (Solanum tuberosum) plants at different distances, using a kanamycin resistnace transgene (nptII) as a selectable marker. All potato plants were from the variety Désirée. Non-transgenic potato plants, used as potential recipients of transgene-containing pollen, were planted in 12 sub-plots, at distances of 0–20 m from the nearest transgenic potato plants. Seeds harvested from the non-transgenic plants were screened for resistance to kanamycin, and molecular methods were used to confirm that resistant progeny contained thenptII gene. Where transgenic and non-transgenic potato plants were in alternate rows (leaves touching), 24% of seedlings from the non-transgenic parent plants were kanamycin-resistant. Comparable seedlings from plants at up to 3 m distance had a resistance frequency of 2%, at 10 m the frequency was 0.017% and at 20 m no resistant progeny were observed. Plants of the weed speciesS. dulcamara andS. nigrum were also planted close to the transgenic potatoes to test for evidence of hybridization, and no kanamycin-resistant seedlings were observed among progeny fromS. dulcamara andS. nigrum. This investigation provided evidence that the extent of gene dispersal from transgenic potatoes to non-transgenic potatoes falls markedly with increasing distance, and is negligible at 10 m. There was, also, no evidence of transgene movement from potato toS. dulcamara andS. nigrum under field conditions. These data will be valuable in defining genetic isolation procedures for the early field evaluation and the use of novel transgenic potato genotypes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02336774
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  • 7
    Electronic Resource
    Electronic Resource
    Opportunities for gene transfer from transgenic oilseed rape (Brassica napus) to related species (1994)
    Springer
    Transgenic research 3 (1994), S. 263-278 
    Details
    ISSN: 1573-9368
    Keywords: Brassica napus ; oilseed rape ; transgenic plants ; interspecific hybridization ; gene transfer ; risk assessment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Before novel transgenic plant genotypes are grown outside containment facilities and evaluated under field conditions, it is necessary to complete a risk assessment to consider the possible consequences of that release. An important aspect of risk assessment is to consider the likelihood and consequences of the transgene being transferred by cross-pollination to related species, including other crops, weeds and ruderal populations. The purpose of this report is to review the literature to assess the ease with whichBrassica napus can hybridize with related species. The evidence for hybridization is considered at three levels: a) by open pollination, b) by hand pollination and c) by the use ofin vitro ovule and embryo rescue techniques; and also examines the fertility and vigour of the F1, F2 and backcross generations. Four species are reported to hybridize withB. napus by open pollination:B. rapa andB. juncea using fully fertile parents; andB. adpressa andR. raphanistrum using a male-sterileB. napus parent. Seventeen species are reported to form hybrids (including the four species above) withB. napus when pollination is carried out manually. At least 12 of these species were unable to form F2 progeny, and eight were unable to produce progeny when the F1 was backcrossed to one of the parental species. Many factors will influence the success of hybridization under field conditions, including: distance between the parents, synchrony of flowering, method of pollen spread, specific parental genotypes used, direction of the cross and the environmental conditions. Even where there is a possibility of hybridization betweenB. napus and a related species growing in the vicinity of a release, poor vigour and high sterility in the hybrids will generally mean that hybrids and their progeny will not survive in either an agricultural or natural habitat.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01973586
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  • 8
    Electronic Resource
    Electronic Resource
    Liver, renal and subcutaneous histopathology in PEPCK-bGH transgenic pigs (1994)
    Springer
    Transgenic research 3 (1994), S. 401-405 
    Details
    ISSN: 1573-9368
    Keywords: gene transfer ; transgenic ; swine ; growth hormone ; PEPCK-bGH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic pigs were created that harboured a phosphoenol pyruvate carboxykinase-bovine growth hormone construct (PEPCK-bGH). Four founder animals and two transgenic offspring from one line were evaluated between 61/2 and 12 months of age. There was no evidence of severe hepatic or renal lesions in these pigs, which characterised transgenic PEPCK-bGH mice previously described. While glomerular and tubular lesions in kidney sections were not identified in the transgenic pigs, mesangial cell proliferation was observed in two transgenic offspring from a single line. Additionally, glomerular size was significantly increased in four of four puberal transgenic swine when compared to age- and sex-matched controls (28.30±4.1 vs. 14.2±2.7×105 μm3; representing 3 transgenic lines,p〈0.05). Surprisingly, no mature adipocytes were observed in subcutaneous sections obtained in transgenic GH pigs. Histological evaluation of these transgenic pigs further illustrates the requirement for precise control of growth-related genes and their protein products.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01976771
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  • 9
    Electronic Resource
    Electronic Resource
    Agrobacterium rhizogenes—transformed plant roots as a source of grapevine viruses for purification (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 291-301 
    Details
    ISSN: 1573-5044
    Keywords: clesteroviruses ; cytopathology ; fleek disease ; gene transfer ; tissue culture ; virus purification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Agrobacterium rhizogenes-mediated transformation was applied toVitis spp. andNicotiana spp. infected by different grapevine phloem-limited viruses (grapevine fleck virus, grapevine virus A, grapevine virus B) to obtain root cultures for virus purification. All plant species were successfully transformed, and several clones were established in liquid culture. Transformed grapevine roots contained as much virus as non transformed roots and more than leaves, as assessed by ELISA and thin sectioning. Likewise, transformed roots ofNicotiana benthamiana Domin. contained in average more GVA than leaves, especially those at the base and the top of the plant, whereas withNicotiana occidentalis wheel., GVB was apparently less concentrated than in leaves.Nicotiana root grew faster than those ofVitis. All viruses multiplied and persisted in root cultures, which were successfully used for purification. Virus yields were the same (GFkV and GVB) or higher (GVB) than those reported in the literature. Grapevine roots may prove useful for culturing and purifying other non-mechanically transmissible grapevine viruses.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00046086
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  • 10
    Electronic Resource
    Electronic Resource
    Recovery of transgenic trees after electroporation of poplar protoplasts (1994)
    Springer
    Transgenic research 3 (1994), S. 13-19 
    Details
    ISSN: 1573-9368
    Keywords: gene transfer ; electroporation ; stable transformation ; protoplasts ; transgenic trees ; Populus tremula x P. alba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts from leaflets ofin vitro cuttings were electroporated in osmotically adjusted and buffered solutions containing plasmid DNA: pABD1, carrying thenptII gene for resistance to neomycin; pGH1, carrying a mutant acetolactate synthase gene,als, for resistance to sulfonylurea; and pGSFR781A, carrying a synthetic phosphinothricin acetyltransferase (pat) for resistance to phosphinothricin (Basta). Gene transfer was repeatedly efficient, without use of carrier DNA, in the range of one transformant for 105 to 104 protoplast-derived cell colonies. This was probably due to the high plating efficiency (30%) of protoplasts in our culture process. Selection for expression of foreign genes was applied in liquid medium and repeatedly achieved with 30 μM paromomycin for NPTII, 200 nM chlorsulfuron for the mutant ALS ofArabidopsis and 25 μM phosphinothricin for PAT expression. Integration of foreign genes into genomic DNA of resistant poplar trees was demonstrated by Southern blot hybridizations, which revealed that for some transformants practically no other part of the vector plasmid than the selected gene was integrated. Effective processes for protoplast culture, efficient selection at the cell colony stage and gene transfer will provide new possibilities in poplar breeding.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01976022
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  • 11
    Electronic Resource
    Electronic Resource
    Accumulation of type I fish antifreeze protein in transgenic tobacco is cold-specific (1993)
    Springer
    Plant molecular biology 23 (1993), S. 377-385 
    Details
    ISSN: 1573-5028
    Keywords: antifreeze proteins ; gene transfer ; preproprotein processing ; α-helix stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Expression of fish antifreeze protein (AFP) genes in plants is a possible means of increasing their frost resistance and freeze tolerance. Initial work involved transfer into tobacco of an AFP gene from winter flounder which codes for the alanine-rich, α-helical Type I AFP. Plants were transformed with a gene construct in which the preproAFP cDNA was inserted between the cauliflower mosaic virus 19S RNA promoter and the nopaline synthetase polyadenylation site. Although transgenic plants produced AFP mRNA, no AFP was detected on western blots. Re-evaluation of AFP expression in these transgenic plants showed that AFP accumulated to detectable levels only after exposure of the plant to cold. Extracts of plants incubated at 4°C for 24 h contained a protein which co-migrated with winter flounder proAFP and was cross-reactive to Type I AFP antisera. Two other minor protein bands of slightly higher apparent M r also cross-reacted with the antisera and are thought to represent processing intermediates. The proAFP was unique to the transgenic plants and was absent in extracts taken prior to cold exposure. AFP levels increased over the first 48 h of cold incubation then remained stable. Since the α-helix content of Type I AFP has been shown to decrease markedly at warmer temperatures, we postulate that Type I AFP stability in transgenic plants is dependent on its secondary structure.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00029012
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  • 12
    Electronic Resource
    Electronic Resource
    Highly efficient transfer and stable integration of foreign DNA into partially digested rice cells using a pulsed electrophoretic drive (1993)
    Springer
    Transgenic research 2 (1993), S. 245-251 
    Details
    ISSN: 1573-9368
    Keywords: rice ; small cell groups ; electrophoresis ; gene transfer ; stable integration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A new method has been developed to introduce foreign DNA into rice cells. Gene delivery occurred when an electrophoretic drive with cycles of intervallic electric field was applied to a mixture containing partially digested small cell groups (SCGs) and plasmid DNAs. Gene transfer efficiency was evaluated by the detection of β-glucuronidase (GUS) activity resulting from expression of a chimaeric plasmid DNA. The optimal combination of treatment conditions (3 V/cm, 30 s pulse and 30 min electrophoretic run) produced a frequency of up to 8.2% of blue cells in transformed microcalluses 40 days after culture of treated SCGs without selection for kanamycin resistance. Southern hybridization showed that the foreign gene had integrated into the chromosomal DNA. These results demonstrate that pulsed electrophoretic drive is applicable to the transfer of foreign genes into plant cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01968837
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  • 13
    Electronic Resource
    Electronic Resource
    Transgenesis in chickens (1993)
    Springer
    Transgenic research 2 (1993), S. 125-133 
    Details
    ISSN: 1573-9368
    Keywords: chicken embryo ; gene transfer ; retroviral vector ; cloned DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The application of transgenic technology to domestic poultry offers an alternative means to conventional practice for improvement of this highly productive agricultural species. The hen's reproductive system has unique characteristics which have imposed limitations on the use of established methods for artificial gene transfer. In this article, we review the various strategies that have been adopted to overcome the problem. Target sites for gene insertion include the fertilized ovum, the blastodermal embryo in the unincubated egg, and the primordial germ cells. Notable success in obtaining somatic and germline transformation has been achieved with the use of retroviral vectors to infect the blastodermal embryo. Current attempts to introduce DNA directly into the genome, without resort to pathogen-derived vectors, are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01972605
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  • 14
    Electronic Resource
    Electronic Resource
    Development of an airgun device for particle bombardment (1993)
    Springer
    Plant cell, tissue and organ culture 33 (1993), S. 247-250 
    Details
    ISSN: 1573-5044
    Keywords: gene transfer ; GUS gene-marker ; particle acceleration ; transient expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Construction and operation of an airgun device for transient gene-expression studies in monocots is described. Compressed air in a cylinder of an airgun was used as the source of propulsion for DNA-coated gold or tungsten particles. Under a partial vacuum of 700 mm Hg, velocity of the macrocarrier was measured at 520 m sec−1 and 432 m sec−1 at atmospheric pressure. Optimum distance from the stopping plate to different target cells during bombardment ranged from 4 to 7 cm. Mean transformation efficiency of the GUS-gene marker was estimated at 350 transformants per 65 mg fresh weight of the maize cultured cells. Up to 200 GUS transformed cells were detected per 100 mg of embryogenic rice callus. Use of gold flakes or tungsten powder as microcarriers resulted in similar transformation rates. No transformation was observed in any cells when DNA constructs contained prokaryotic translation initiation sequences for the GUS gene. Based on transient GUS assays, further modification of the airgun device will likely be necessary to obtain high stable transformation rates.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02319008
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  • 15
    Electronic Resource
    Electronic Resource
    Herpesvirus-mediated gene delivery into the rat brain: specificity and efficiency of the neuron-specific enolase promoter (1993)
    Springer
    Cellular and molecular neurobiology 13 (1993), S. 503-515 
    Details
    ISSN: 1573-6830
    Keywords: herpesvirus vector ; gene transfer ; neuron-specific enolase (NSE) promoter ; lacZ, stereotactic delivery ; rat CNS
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. Herpesvirus infection with genetically engineered vectors is a way to deliver foreign gene products to various cell populations in culture andin vivo. Selective neuronal gene expression can be achieved using the neuron-specific enolase (NSE) promoter regulating expression of a transgene placed in and delivered by a herpesvirus vector. 2. We sought to determine the anatomical specificity and efficiency of herpesvirus-mediated gene transfer into the rat brain following placement of virus particles carrying a transgene (lacZ) under control of the NSE promoter. The virus utilized was thymidine kinase (TK) deficient and therefore replication deficient in the brain. 3. Infusion of 106 plaque-forming units of virus into the striatum caused a limited number of striatal neurons to express thelacZ transgene mRNA and protein product 7 days postinfection. In addition, small numbers of neurons expressing the transgene mRNA and protein were found ipsilateral to the viral injection in the frontal cortex, substantia nigra pars compacta, and thalamus. Neurons at these anatomic loci project directly to the striatal injection site. No other cells within the brains of injected animals expressed thelacZ gene. 4. While this herpesvirus NSE vector was capable of introducing novel functional genetic information into postmitotic neurons within defined neuroanatomic constraints, the numbers of neurons expressing detectable levels ofβ-galactosidase was minimal. The calculated efficiency of delivery and transgene expression at 7 days postinfection was 1 transgenic neuron per 104 virus particles infused. 5. We conclude that NSE probably is not an optimal promoter for use in gene delivery to CNS neurons in herpesvirus vectors and that the efficacy of gene delivery using other neuron-specific promoters placed at various sites in the herpes viral genome needs to be explored.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00711459
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  • 16
    Electronic Resource
    Electronic Resource
    Gene transfer by protoplast fusion: Expression cloning of rare gene products in mammalian cells (1993)
    Springer
    Methods in cell science 15 (1993), S. 69-71 
    Details
    ISSN: 1573-0603
    Keywords: protoplast fusion ; gene transfer ; alkaline phosphatase ; expression cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A method is described that facilitates performing a large number of protoplast fusions to mammalian cells simultaneously and successfully. This method makes it possible to circumvent typical hurdles to the use of transient expression in mammalian cells, facilitating expression cloning of DNA enoding the newly detected gene product of interest. The original in vitro assay used to define the new activity is of interest, adapted to microtiter plates, combined with protoplast fusion, extends the reach of expression cloning to such cases as products of a rare message, or activities involving a multisubunit or unstable protein or both.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01667364
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  • 17
    Electronic Resource
    Electronic Resource
    Potential for wild species in cool season food legume breeding (1993)
    Springer
    Euphytica 73 (1993), S. 109-114 
    Details
    ISSN: 1573-5060
    Keywords: wild-germplasm ; interspecific-hybridization ; gene pools ; introgression ; gene transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Wild species which are crossable to cultivated pea, lentil, and chickpea have been collected and are maintained in major germplasm collections throughout the world. Wild species of Vicia crossable to the cultivated faba bean have not been found. The primary, secondary, and tertiary gene pools of the cool season food legumes represent potential genetic diversity that may eventually be exploited in cultivated types to overcome biotic and abiotic stresses. Technical difficulties in obtaining hybrids beyond those within the primary gene pool is a major obstacle. Reproductive isolation, embryo breakdown, hybrid sterility, and limited genetic recombination are major barriers to greater use of wild germplasm. Conventional crossing has been successful in producing interspecific hybrids in Lens, Cicer and Pisum and those hybrids are being evaluated for desired recombinants. In vitro culture of hybrid embryos has been successful in overcoming barriers to wider crosses in Lens. The successful transfer of genes from wide sources to cultivated types can be assisted by repeated backcrossing and selection designed to leave behind undesired traits while transferring genes of interest. Molecular marker assisted selection may become a valuable tool in the future use of wild species. In general, too little is known about the possible genetic variation available in wild species that could be valuable in developing resistance to biotic and abiotic stresses. Current efforts on the use of wide hybridization in the cool season food legumes are reviewed and discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027187
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  • 18
    Electronic Resource
    Electronic Resource
    The potential of gene technology and genome analysis for cool season food legume crops: theory and practice (1993)
    Springer
    Euphytica 73 (1993), S. 177-189 
    Details
    ISSN: 1573-5060
    Keywords: gene cloning ; gene transfer ; virus and insect resistance ; genome analysis ; DNA figerprinting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The potential of plant gene technology encompasses a multitude of different techniques ranging from the isolation of useful genes, their characterization and in vitro manipulation to the reintroduction of the modified constructs into target plants, where they are expressed at a rate that alters the phenotype of the plants. Genome analysis, on the other hand, aims at characterizing the genome architecture and function(s). Plant gene technology has catalyzed progress in plant breeding, as will be exemplified by a few examples, but has not yet been applied to food legume improvement on a large scale. Genome analysis, however, has a series of practical implications, as is illustrated by the successful introduction of DNA fingerprint and PCR fingerprint techniques to chickpea (Cicer arietinum L.) breeding and Ascochyta rabiei pathotyping. The present overview addresses both areas of plant molecular biology to illustrate their potential for food legume breeding.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027193
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  • 19
    Electronic Resource
    Electronic Resource
    Particle bombardment-mediated transient expression of a Brazil nut methionine-rich albumin in bean (Phaseolus vulgaris L.) (1992)
    Springer
    Plant molecular biology 20 (1992), S. 357-359 
    Details
    ISSN: 1573-5028
    Keywords: Brazil nut ; 2S albumin gene ; gene transfer ; Phaseolus vulgaris ; particle bombardment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Bean (Phaseolus vulgaris L.) mature embryos were transformed using biolistic methods with a plasmid containing 2S albumin and β-glucuronidase structural sequences, both under the control of the 35S CaMV promoter. We have shown that chimaeric tissues could be obtained and that both structural sequences were expressed to similar levels.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014508
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  • 20
    Electronic Resource
    Electronic Resource
    Factors influencing Agrobacterium tumefaciens mediated transformation and expression of kanamycin resistance in pickling cucumber (1992)
    Springer
    Plant cell, tissue and organ culture 31 (1992), S. 185-193 
    Details
    ISSN: 1573-5044
    Keywords: Agrobacterium transformation ; Cucumis sativus ; gene transfer ; neomycin phosphotransferase ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cucumber (Cucumis sativus L.) petiole and leaf segments of two pickling genotypes were transformed with Agrobacterium tumefaciens strain LBA 4404, an octopine Ti-plasmid deletion mutant that is avirulent (disarmed plasmid), but to which were added T-DNA inserts on binary plasmids (pBIN 19, ca. 10 kb, and pCGN 783, ca. 25 kb). Expression of neomycin phosphotransferase (NPT II) encoding resistance to the aminoglycoside kanamycin was used as a selectable marker. Factors which influenced the frequency of callus development on medium containing kanamycin (75 mg l-1) were explant size, bacterial concentration and length of exposure, cocultivation period, and presence of acetosyringone. The optimal procedure involved exposing segments of petiole (4–6 mm) or leaf (0.5 cm2) segments to a bacterial suspension (108 cells ml-1) containing 20 μM acetosyringone for 5 min, followed by a 48 h cocultivation period on a tobacco feeder layer. Explants were placed on MS medium containing 500 mg l-1 carbenicillin, 75 mg l-1 kanamycin, and NAA/BA (5.0/2.5 μM) or 2,4-d/BA (5.0/5.0 μM) and subcultured twice, each after a 2–3 week period, onto fresh media. The overall frequency of transformed callus was 20–50%; the frequency of plantlet regeneration from transformed callus was 8–15%. Twenty-one out of 23 individual plants recovered from two genotypes of pickling cucumber were NPT II positive (transformation frequency of 9%). Copy number of the NPT II gene insert (35S-NPT II-3′ fragment, ca. 2.2 kb) in three transformed plants was estimated at ten per haploid genome, indicative of multiple insertions within the cucumber genome. Multimers of the gene (visible as 4.4 and 6.6 kb fragments in Southern analysis) were detected in one plant, suggestive of tandem duplications or repeats. Progeny from a cross between this transformed plant and a nontransformed control showed segregation for the NPT II gene in dot-blot assays; at least 24 plants out of 32 were kanamycin positive. Copy number in the progeny was variable, and ranged from none to ten.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00036222
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  • 21
    Electronic Resource
    Electronic Resource
    Differential binding of rat pituitary-specific nuclear factors to the 5′-flanking region of pituitary and placental members of the human growth hormone gene family (1991)
    Springer
    Molecular and cellular biochemistry 106 (1991), S. 181-187 
    Details
    ISSN: 1573-4919
    Keywords: human growth hormone ; rat pituitary tumor cells ; tissue-specificity ; gene transfer ; DNase 1 footprinting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary Placental chorionic somatomammotropin (hCS-A or B) and growth hormone variant (hGH-V) are members of the human growth hormone family, and are related by structure and function to pituitary growth hormone (hGH-N). However, while the hGH-N gene is expressed specifically in the anterior pituitary, hGH-V and hCS are produced in the placenta. Hybrid hGH-N, hGH-V and hCS-A genes containing 5′-flanking sequences, including the endogenous promoter, are preferentially expressed in rat pituitary tumor (GC) cells, after gene transfer. Since interaction with a pituitary-specific protein (Pit 1) is required for efficient hGH-N as well as rat growth hormone (rGH) gene expression in GC cells, binding of pituitary proteins to the hGH-V and hCS-A promoter sequences was investigated. Rat Pit 1 binds at two locations on the hGH-N gene, a distal (−140/−107) and proximal site (−97/−66), in a similar manner to that observed with the rGH gene. By contrast, efficient Pit 1 binding was seen only to the distal site of the hGH-V gene and the proximal site of the hCS-A gene. Although binding of a protein to the distal hCS-A sequences was observed, the site of interaction was truncated (−140/−116), not pituitary-specific, and was more consistent with the binding of Sp1. These data indicate that rat Pit 1 binds to the placental hGH-V and hCS-A genes and correlates with their promoter activity in GC cells after gene transfer. However, the data also indicate that rat Pit 1 binds to human and rat pituitary growth hormone in a similar manner (two sites of interaction) and that the pattern of binding is distinct from the placental members of the hGH gene family. These data indicate that human Pit 1, unlike the rat equivalent, might distinguish these genes functionally (tissue-specifically) as well as structurally.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00230184
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  • 22
    Electronic Resource
    Electronic Resource
    Genetic improvement of legumes using somatic cell and molecular techniques (1991)
    Springer
    Euphytica 55 (1991), S. 157-169 
    Details
    ISSN: 1573-5060
    Keywords: gene transfer ; genetic manipulation ; chimaeric genes ; legumes ; transformation ; somatic hybridisation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The merits and limitations of somatic cell techniques involving Agrobacterium-mediated transformation, direct gene transfer and protoplast fusion, are discussed in relation to the genetic improvement of forage and grain legumes. Whilst progress with legumes is limited compared to that with plants of other families such as the Solanaceae, the fact that many legumes are readily amenable to tissue culture now permits somatic cell techniques to be targetted to these species. Future development of the subject will necessitate close collaboration between molecular biologists and plant breeders to enable novel plants generated by in vitro technologies to be incorporated into conventional breeding programmes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00025229
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  • 23
    Electronic Resource
    Electronic Resource
    Hereditary human myopathies in muscle culture (1991)
    Springer
    Neurological sciences 12 (1991), S. 257-268 
    Details
    ISSN: 1590-3478
    Keywords: Human muscle cultures ; clones ; nerve-muscle cocultures ; cell lines ; heterokaryons ; myoblast transfer ; gene transfer ; Myo D ; cybrid clones ; hereditary human myopathies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Sommario L'Autore illustra l'utilità e i limiti delle colture di muscolo umano nello studio delle miopatie umane ereditarie. In particolare nelle miopatie dovute a difetti di enzimi-specifici muscolari, l'utilizzo di tecniche di cocoltura muscolo-nervo con avanzato grado di maturazione muscolare permette di delucidare i meccanismi molecolari patogenetici di tali malattie. L'uso di avanzate tecniche di biologia molecolare e cellulare, quali linee permanenti, trapianto di mioblasti, trasferimento genico e di mitocondri, oltre che fornire utili informazioni a livello molecolare potranno trovare applicazione, in futuro, persino nella terapia di molte miopatie ereditarie.
    Notes: Abstract In this article I illustrated the use of regenerating human muscle cultures for studying the hereditary human myopathies. Although some of the data are still controversial, they do point up the great potential of this “in vitro system”. For hereditary myopathies due to developmentally regulated proteins that are expressed only at a more advanced stage of muscle differentiation, the use of highly differentiated nerve-muscle cocultures might contribute significantly to a better understanding of their developmental pathogenesis. More advanced techniques (permanent human muscle cell lines, heterokaryons, myoblast transfer, gene transfer, myogenic conversion of human non-muscle cells, cybrid clones) may provide a great deal of information at molecular level and may also have practical applications in the diagnosis or even in the treatment of hereditary human myopathies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02337773
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  • 24
    Electronic Resource
    Electronic Resource
    Long-term expression of gene introduction into normal human T-lymphocytes by retroviral-mediated gene transfer (1991)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 45 (1991), S. 353-358 
    Details
    ISSN: 0730-2312
    Keywords: bone marrow ; peripheral blood ; gene transfer ; IL-2 ; neo gene ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Human T-lymphocytes are long lived, easily accessible, mature, and capable of proliferation. They are theoretically a suitable target for retroviral mediated gene transfer. To test this hypothesis, normal human T-cells obtained from bone marrow and peripheral blood were stimulated with phytohemagglutinin (PHA) and infected 24 h later with the retroviral vector N2 which carries the bacterial neo gene. T-lymphocytes were propagated in culture for up to 14 weeks with interleukin-2 (IL-2). Analysis by whole cell RNA dot/blot using a single stranded RNA probe demonstrated persistent expression of the neo gene. Preliminary functional studies revealed that both helper and suppressor functions were preserved in the infected cells in culture. These results demonstrate that normal T-cells are capable of long-term expression of genes introduced by retroviral mediated gene transfer and are potential target cells for somatic gene therapy.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240450408
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  • 25
    Electronic Resource
    Electronic Resource
    Shoot apical meristems as a target for gene transfer by microballistics (1955)
    Springer
    Euphytica 85 (1955), S. 45-51 
    Details
    ISSN: 1573-5060
    Keywords: meristem ; shoot apex ; ballistic microtargeting ; gene transfer ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The classical approach of gene transfer to a given plant species delivers the foreign gene to transformable cells and then puts the effort into generating plants. This approach is very difficult in many important crop plants, including cereals, and the results of regeneration are very genotype-dependent. In contrast, we use regenerable cells and try to transform them. Shoot apical meristems provide a tissue which regenerates in situ a fertile plant for most given genotypes or species. Transformation of meristem cells may lead to transgenic sectors in chimeras. These sectors may contribute to the gametes and, thus, to transgenic offspring, which then should be homohistonts and not sectorial chimeras like their parents. Our model plant for these studies is wheat. Microtargeting is a ballistic approach which is particularly suitable for the controlled delivery of microprojectiles to meristem cells in situ (Sautter et al., 1991). We summarize in this paper our experience with ballistic microtargeting of transgenes to wheat shoot apical meristem cells in situ.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023929
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  • 26
    Electronic Resource
    Electronic Resource
    Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants (1955)
    Springer
    Euphytica 85 (1955), S. 81-88 
    Details
    ISSN: 1573-5060
    Keywords: gene transfer ; Hordeum vulgare ; neomycin phosphotransferase II ; particle bombardment ; transgenic barley
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Transgenic barley plants (Hordeum vulgare L. cv. Kymppi) were obtained by particle bombardment of various tissues. Immature embryos and microspore-derived cultures were bombarded with gold particles coated with plasmid DNA carrying the gene coding for neomycin phosphotransferase II (NPTII), together with plasmid DNA containing the gene for β-glucuronidase (GUS). Bombarded immature embryos were grown to plants without selection and NPTII activity was screened in small plantlets. One plant proved to be transgenic (T0). This chimeric plant passed the transferred nptII gene to its T1 progeny. The presence of the nptII gene was demonstrated by the PCR technique and enzyme activity was analyzed by an NPTII gel assay. Four T0 spikes and 15 T1 offspring were transgenic. The integration and inheritance was confirmed by Southern blot hybridization. Transgenic T2 and T3 plants were produced by isolating embryos from green grains of transgenic T1 and T2 plants, respectively and growing them to plants. After selfing, the ratio of transgenic to non-transgenic T2 offspring was shown to follow the rule of Mendelian inheritance. The general performance of transgenic plants was normal and no reduction in fertility was observed. Microspore-derived cultures were bombarded one and four weeks after microspore isolation. After bombardment, cultures were grown either with or without antibiotic selection (geneticin R or kanamycin). When cultures were grown without selection and regenerated plants were transferred to kanamycin selection in rooting phase, one out of a total of about 1500 plants survived. This plant both carried and expressed the transferred nptII gene. The integration was confirmed by Southern blot hybridization. This plant was not fertile.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023933
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  • 27
    Electronic Resource
    Electronic Resource
    The sulphur-rich Brazil nut 2S albumin is specifically formed in transgenic seeds of the grain legume Vicia narbonensis (1955)
    Springer
    Euphytica 85 (1955), S. 181-192 
    Details
    ISSN: 1573-5060
    Keywords: Vicia narbonensis ; gene transfer ; gene expression ; seeds ; 2S albumin ; methionine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Epicotyl explants were co-cultivated with Agrobacterium tumefaciens EHA101 to transfer a chimeric 2S albumin gene construct carried in the binary Ti plasmid vectors pGSGLUC1 or pGA472 into the grain legume Vicia narbonensis. This gene encoding the sulphur-rich Brazil nut albumin was under the control of either the CaMV 35S promoter which permits gene expression in all organs, or the Vicia faba legumin B4 promoter which elicits seed-specific gene expression. After callus formation and selection for kanamycin resistance, somatic embryos were induced which, in the case of transformation with the vector pGSGLUC1, were screened for GUS activity. Embryos that produced GUS were in addition analysed for 2S albumin formation. Selected transgenic embryos were cloned by multiple shoot regeneration. Rooted and fertile plants were obtained by grafting transgenic shoots on the appropriate seedlings. R1 and R2 generations were raised and analysed for GUS as well as 2S albumin gene expression. Expression of the 35S promoter/2S albumin gene fusion took place in all organs of the transgenic plants including the cotyledons of seeds, whereas seed-specific gene expression was found in transformants with the legumin promoter/2S albumin gene fusion. The 2S albumin accumulated in the 2S protein fraction of transgenic seeds and its primary translation product was processed into the 9 and 3 kDa polypeptide chains. The foreign protein was localised in the protein bodies of the grain legume. Analysis of the R2 plants indicated Mendelian inheritance of the 2S albumin gene. In homozygous V. narbonensis plants the amounts of 2S albumin were twice that present in the corresponding heterozygous plants. Whereas only low level formation of the foreign protein was achieved if the gene was under the control of the 35S promoter, approximately 3.0% of the soluble seed protein was 2S albumin if seed-specific gene expression was directed by the legumin B4 promoter. Some of these transformants exhibited a three-fold increase in the methionine content of the salt-soluble protein fraction extracted from seeds.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023947
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  • 28
    Electronic Resource
    Electronic Resource
    Strategies for variety-independent genetic transformation of important cereals, legumes and woody species utilizing particle bombardment (1955)
    Springer
    Euphytica 85 (1955), S. 13-27 
    Details
    ISSN: 1573-5060
    Keywords: gene transfer ; crop species ; particle bombardment ; transgenic plants ; cereals ; legumes ; woody plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The limiting component in the creation of transgenic crops has been the lack of effective means to introduce foreign genes into elite germplasm. However, the development of novel direct DNA transfer methodology, by-passing limitations imposed by Agrobacterium-host specificity and cell culture constraints, has allowed the engineering of almost all major crops, including formerly recalcitrant cereals, legumes and woody species. The creation of transgenic rice, wheat, maize, barley, oat, soybean, phaseolus, peanut, poplar, spruce, cotton and others, in an efficient and in some cases, variety-independent fashion, is a significant step towards the routine application of recombinant DNA methodology to the improvement of most important agronomic crops. In this review we will focus on key elements and advantages of particle bombardment technology in order to evaluate its impact on the accelerated commercialization of products based on agricultural biotechnology and its utility in studying basic plant developmental processes and function through transgenesis. Fundamental differences between conventional gene transfer methods, utilizing Agrobacterium vectors or protoplast/suspension cultures, and particle bombardment will be discussed in depth.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023926
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