A vehicle for the introduction of transposons into plant-associated pseudomonads

doi:10.1016/0147-619X(85)90043-5

Plasmid

Volume 13, Issue 3, May 1985, Pages 200-204

https://doi.org/10.1016/0147-619X(85)90043-5 Get rights and content

Abstract

A recombinant plasmid with wide-host-range transfer functions, narrow-host-range replication functions, and carrying a kanamycin-resistant transposon transferred kanamycin resistance to a number of plant-associated pseudomonads. Southern hybridization studies suggest that only a small portion of the plasmid, coinciding with the location of the transposon, is present in the kanamycin-resistant Pseudomonas derivatives. The plasmid sequences appear to be inserted at a number of different sites in the recipient genome. This plasmid can thus be used as a vehicle for the introduction of transposons into some plant-associated pseudomonads and should be useful in both genetic and ecological studies of these bacteria.

References (17)

N. Kleckner et al.
J. Mol. Biol
(1977)
D.F. Myers et al.
Trans. Brit. Mycol. Soc
(1983)
A. Oka et al.
J. Mol. Biol
(1981)
M. Sato et al.
Plasmid
(1981)
R.W. Davis et al.
Advanced Bacterial Genetics
(1980)
D.H. Figurski et al.
N.D.F. Grindley et al.
T. Leisinger et al.
Microbiol. Rev
(1979)

There are more references available in the full text version of this article.

Cited by (17)

Improvement of ectoine productivity by using sugar transporter-overexpressing Halomonas elongata
2016, Enzyme and Microbial Technology
Citation Excerpt :
The primers used in this study are listed in Table S1. All plasmids were introduced into H. elongata using E. coli HB101/pRK2013-mediated conjugation [23]. To eliminate E. coli cells after plasmid transfer, a streptomycin-resistant strain of H. elongata was isolated by serial plating on LB medium containing streptomycin (500 mg/L) and 6% NaCl with incubation at 37 °C.
We successfully enhanced the productivity of ectoine with Halomonas elongata by improvement of the transport of sugar. First, we carried out screening for sugar transporters capable of improving glucose and xylose consumption. We found two transporters: b3657 from Escherichia coli, which is capable of improving glucose consumption, and HEO_0208 from H. elongata, which is capable of improving xylose consumption. Using transporter-overexpressing strains, the productivity of ectoine was improved. These results indicate that sugar consumption is important for efficient ectoine production. As result of phenotypic analysis of a HEO_0208 deletion strain, we discovered that HEO_0208 is the major xylose transporter in H. elongata. This is the first report demonstrating improvement of ectoine productivity by enhancing the transport of sugar.
Ectoine production from lignocellulosic biomass-derived sugars by engineered Halomonas elongata
2013, Bioresource Technology
Citation Excerpt :
The amplified fragments were digested and ligated into the SpeI/NsiI restriction sites of pHS15N. The resulting plasmid, designated pHS15N–lysC, was transferred into H. elongata using Escherichia coli HB101/pRK2013-mediated conjugation (Lam et al., 1985). To eliminate E. coli cells after plasmid transfer, a spontaneous streptomycin-resistant mutant of H. elongata was isolated by serial plating on LB medium containing streptomycin (60 mg/L) and 6% NaCl and incubation at 37 °C.
In this study, the water-retaining cyclic amino acid ectoine was produced from a variety of sugars, including glucose, xylose, cellobiose, and glucose/xylose mixture using engineered Halomonas elongata. When grown on xylose as the sole carbon source, H. elongata produced 333 mmol/kg fresh cell weight (FW) of ectoine, which was 1.4-fold higher than that produced from glucose. To improve ectoine production, an ectD deficient H. elongata mutant was constructed. The engineered H. elongata produced 377 mmol/kg FW of ectoine from a glucose/xylose mixture. Ectoine was also produced from rice straw hydrolysate. These results show that H. elongata can produce ectoine from a variety of sugars derived from lignocellulosic biomass and thus has tremendous potential as a host for producing useful compounds from biomass resources.
Freezing tolerant tobacco, transformed to accumulate osmoprotectants
2002, Plant Science
Abiotic stresses are the major environmental challenges for plant growth and crop productivity. To cope with them various adaptation strategies have been developed by plants, including accumulation of osmoprotectants. Tobacco commercial cultivar was transformed to accumulate proline, fructan or glycine betaine. We used constructs harboring arabidopsis- or vigna-derived genes (AtP5Cs, VacP5Cs) for Δ¹-pyroline-5-carboxylate synthetase production, SacB gene coding for levansucrase from Bacillus subtilis or the codA gene coding for choline oxidase from Arthrobacter globiformis. The reactions to sub-zero temperatures and osmotic stress were followed in several subsequent progenies. Stable transgenic lines have been selected. They survive freezing stress at controlled and field conditions. This is, we believe, the first report for freezing tolerance in proline and fructan transformed plants.
Silene plastocyanin is fully functional in transgenic tobacco
1992, Plant Science
Transgenic tobacco plants, expressing the Silene pratensis (white campion) gene for the precursor of plastocyanin, were analysed with respect to the functionality of the gene product. The gene was found to be expressed in all tissues that were examined due to the strong and constitutive cauliflower mosaic virus 35S promoter. In green tissue the Silene protein was transported into chloroplasts and routed to the chloroplasts lumen, where it was found processed to its mature size. In non-photosynthetic tissue the protein is transported into chromoplasts or leucoplasts. In plants grown in tissue culture the amount of endogenous tobacco plastocyanin was found to be reduced significantly, but the Silene plastocyanin was clearly detectable. When plants were dependent on photosynthesis for growth, due to depletion of sucrose from the medium, still only Silene plastocyanin was present. This strongly suggests that the Silene protein can take over photosynthesis in transgenic tobacco when the endogenous plastocyanin is not present. Silene plastocyanin is considered to be fully functional, since both its transport to the chloroplast and its function in photosynthesis were retained in transgenic tobacco plants.
Plasmid promiscuity and chastity and its uses
1990, Biotechnology Advances
Bacterial plasmids are obligate and intracellular genetic elements that replicate and are maintained autonomously from the chromosome. They are ubiquitous. Some of them are relatively more promiscuous than others. Plasmid genetic systems that contribute to relative promiscuity or chastity in naturally occurring plasmids are described and discussed. Both the promiscuity and the chastity of plasmid-based genetic systems have applications in bacterial molecular genetics, in the production of recombinant DNA products and in the breeding and use of desirable bacteria. The role of these systems in such applications is considered
Vectors permitting visual monitoring of simple transposition events
1989, Gene
The construction and use of two novel transposon(Tn)-delivery vectors is described. These vectors carry Inc.W or Inc.N broad-host-range transfer functions cloned next to the narrow-host-range replicon of pBR329. The host specificities of pSLX10 and pSLX23 both complement and extend the host specificities of existing Tn delivery vectors. Plasmids pSLX10 and pSLX23 were shown to transfer at high frequency in intergeneric matings. The lux genes which are present on each vector permit the visual monitoring of transconjugants which have retained a Tn element, but are devoid of plasmid molecules. pSLX10 and pLSX23 were efficiently used to generate a range of auxotrophic mutants in various strains of Pseudomonas as well as to clone genes from Serratia liquefaciens. These vectors may have general applicability to identify and clone genes in a wide range of Gram-negative bacteria.

View all citing articles on Scopus

¹: Present Address: CIBA-GEIGY Biotechnology Research, P.O. Box 12257, Research Triangle Park, N. C. 27709.

View full text

Short communicationA vehicle for the introduction of transposons into plant-associated pseudomonads

Abstract

J. Mol. Biol

Trans. Brit. Mycol. Soc

J. Mol. Biol

Plasmid

Advanced Bacterial Genetics

Microbiol. Rev

Short communication
A vehicle for the introduction of transposons into plant-associated pseudomonads