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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Urological research 27 (1999), S. 83-96 
    ISSN: 1434-0879
    Keywords: Key words Genitourinary malignancies ; Recombinant DNA ; Gene cloning ; DNA chip technology ; Diagnostics ; Gene therapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Cancer, including genitourinary malignancy, is a consequence of accumulated genetic aberrations in genes involved in crucial regulatory pathways. The result is a deregulation of cellular behaviour, leading to neoplastic transformation, uncontrolled cell proliferation and acquisition of metastatic ability. The development and perfection of techniques in the field of recombinant DNA technology, gene cloning, (differential) analysis of gene expression, and sequencing of genes and proteins have provided a wealth of information about the genetic aberrations associated with cancer development. This “recombinant DNA and gene cloning” technology and the recently developed DNA chip technology may provide new molecular diagnostic tools. Furthermore, the technology of gene cloning in combination with the progress in in vivo gene delivery techniques offers new treatment modalities, like gene therapy, additional to conventional therapies. This review is intended to provide a general introduction to the fundamentals or strategies of recombinant DNA and gene cloning techniques as a basis for understanding the rapidly expanding range of new diagnostic and therapeutic opportunities. Some illustrative examples are provided, addressing basic and biomedical research and possible clinical applications in genitourinary oncology.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: Key words Restriction endonuclease ; Methylase selection ; Gene expression ; DNA methylation ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The genes encoding the ApaLI (5′-G^TGCAC-3′), NspI (5′-RCATG^Y-3′), NspHI (5′-RCATG^Y-3′), SacI (5′-GAGCT^C-3′), SapI (5′-GCTCTTCN1^-3′, 5′-^N4GAAGAGC-3′) and ScaI (5′-AGT^ACT-3′) restriction-modification systems have been cloned in E.␣coli. Amino acid sequence comparison of M.ApaLI, M.NspI, M.NspHI, and M.SacI with known methylases indicated that they contain the ten conserved motifs characteristic of C5 cytosine methylases. NspI and NspHI restriction-modification systems are highly homologous in amino acid sequence. The C-termini of the NspI and NlaIII (5′-CATG-3′) restriction endonucleases share significant similarity. 5mC modification of the internal C in a SacI site renders it resistant to SacI digestion. External 5mC modification of a SacI site has no effect on SacI digestion. N4mC modification of the second base in the sequence 5′-GCTCTTC-3′ blocks SapI digestion. N4mC modification of the other cytosines in the SapI site does not affect SapI digestion. N4mC modification of ScaI site blocks ScaI digetion. A DNA invertase homolog was found adjacent to the ApaLI restriction-modification system. A DNA transposase subunit homolog was found upstream of the SapI restriction endonuclease gene.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical medicine and bioethics 17 (1996), S. 279-314 
    ISSN: 1573-1200
    Keywords: Recombinant DNA ; transgenesis ; patents ; human life forms
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Philosophy
    Notes: Abstract The rationale of patents on transgenic organisms leads to the startling notion of the human qua infringement. The moral reasons by which we may tenably reject such notion are not conclusive as to human life forms outside the body. A close look at recombinant DNA experimentation reveals ingenious processes, but not entities that the body lacks. Except for artificial genes, the genes of biotechnology are found on chromosomes, albeit nonconsecutively, and their uninterrupted transcripts appear in messenger RNA. An enhanced form of protection for ingenious processes, the “human methods patent,” is proposed and defended as a replacement for product patents. The proposed patent would pertain to biotechnology manufacturing and genetic intervention in somatic and germ cells. A counterpart could govern nonhuman life forms. It is argued that compulsory licensing protections should be a condition of such patent. Contrary to the conservative assumption that statutory sobriquets suffice, the reckoning of what qualifies as a patentable ingenious process will continue to require systematic scientific guidance.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0983
    Keywords: Recombinant DNA ; Filamentous fungi ; 5-fluoro-orotic acid ; Homologous transformation system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A homologous gene transfer system for Aspergillus awamori for site-specific integration is described, based on two components. First, a defined A. awamori pyrG mutant strain constructed by a selection strategy for gene-replacement in fungi. Second, a vector with a homologous pyrG selection marker containing a defined mutation at a site different from that of the mutations in the pyrG gene of the defined mutant strain. Defined mutation in the A. awamori pyrG gene, isolated from a genomic library by heterologous hybridisation with the A. niger pyrG gene as a probe, were introduced by specifically altering sequences at restriction sites in the coding region of the gene. After transformation of the A. awamori wild-type strain with vectors containing these mutated pyrG genes, and selection for 5-fluoro-orotic acid resistance (5-FOAR), on the average 60% of the 5-FOAR colonies originated from replacement of the wild-type pyrG gene by the mutated pyrG allele. After transformation of a mutant strain, carrying a mutation near the 5′ end of the pyrG gene with vectors containing a mutation near the 3′ end of the pyrG gene, 35% of the resulting transformants contained one copy of the vector at the pyrG locus.
    Type of Medium: Electronic Resource
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