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Expression of cloned genes in eukaryotic cells using vector systems derived from viral replicons

  • Chapter
Book cover Genetic Engineering 3

Part of the book series: Genetic Engineering ((GEPM))

Abstract

The application of recombinant DNA techniques has revolutionized our understanding of the structure, organization and expression of eukaryotic genes. Well-established methodologies are available for the construction of clone libraries containing either cDNA copies of mRNAs (Williams, this series, Vol. 1) or genomic DNA (Dahl et al., this series, Vol. 2) and any sequence can be isolated from such libraries if a method for screening is available. Once a suitable clone has been isolated, even if it contains only a small portion of the gene or genetic locus in question, the entire gene or locus, together with large amounts of flanking DNA, can be isolated by repeated screenings of appropriate libraries (chromosome walking) and the organization of these sequences can be studied in the source organism and in genetically related individuals and species using the transfer hybridization technique developed by Southern (1975). The availability of chemical DNA sequencing techniques (Maxam and Gilbert, 1980) and, more particularly, the development by Sanger and his colleagues of the extraordinarily rapid dideoxy sequencing method which employs cloning in M13 (Sanger, 1981), mean that the determination of the complete nucleotide sequence of large segments of cloned DNA is now routine. Finally, the adaptation of the original transfer hybridization technique to the detection of RNA (Alwine et al., 1977; Thomas, 1980) and the establishment and refinement of the endonuclease SI method for mapping the 5′ and 3′ termini of mRNA molecules and the location of introns (Berk and Sharp, 1977; Favaloro et al., 1980) allow the pattern of transcription to be readily analysed if a cloned probe is available.

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  1. Cancer Research Campaign Eukaryotic Molecular Genetics Research Group, Department of Biochemistry, Imperial College of Science and Technology, London, SW72AZ, UK

    Peter W. J. Rigby

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  1. St Mary’s Hospital Medical School, University of London, UK

    Robert Williamson (Professor of Biochemistry) (Professor of Biochemistry)

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Rigby, P.W.J. (1982). Expression of cloned genes in eukaryotic cells using vector systems derived from viral replicons. In: Williamson, R. (eds) Genetic Engineering 3. Genetic Engineering. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-7078-3_3

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  • DOI: https://doi.org/10.1007/978-1-4615-7078-3_3

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