Abstract
The present methods for isolating a genomic clone for a given human gene are based primarily on the isolation of DNA clones starting with mRNA from an expressing tissue or cell line or a sequence information from the protein that allows synthesis of a nucleotide probe specific enough for screening of DNA libraries. Isolation of genes from genomic DNA by direct expression in a heterologous animal cell line has been achieved for genes that confer a directly selectable phenotype to the heterologous cell line, e.g., genes for thymidine kinase (Perucho et al. 1980a), adenosine phosphotransferase (Lowy et al. 1980), hypoxanthine guanosine phosphotransferase, or transforming genes (Goldfarb et al. 1982, Shih and Weinberg 1982). The reisolation depends on the linkage of marker sequences that can be screened or selected for in E. coli, e.g., human repetitive sequences (Shih and Weinberg 1982) or antibiotic resistance markers (Lund et al. 1982). Normally several rounds (at least two) or transfection as well as preparation and screening of a complete genomic DNA library from the expressing clone(s) is necessary for gene isolation.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1522
Blin N, Stafford DW (1976) Isolation of high-molecular-weigth DNA. Nucleic Acids Res 3:2303–2308
Bradshaw HD (1983) Molecular cloning and cell cycle-specific regulation of a functional human thymidine kinase gene. Proc Natl Acad Sci USSA 80:5588–5591
Cami B, Kourilsky P (1978) Screening of cloned recombinant DNA in bacteria by in situ colony hybridization. Nucleic Acids Res 5:2381–2390
De Saint Vincent BR, Delbrück S, Eckhart W, Meinkoth J, Vitto L, Wahl G (1981) The cloning and reintroduction into animal cells of a functional CAD gene, a dominant amplifiable genetic marker. Cell 27:267–277
Goldfarb M, Shimizu K, Perucho M, Wigler M (1982) Isolation and preliminary characterization of a human transforming gene from T24 bladder carcima cells. Nature (Lond) 296:404–409
Goodenow RS, McMillan M, Örn A, Nicolson M, Davidson N, Frelinger JA, Hood L (1982) Identification of a Balb/c H-2L gene by DNA mediated gene transfer. Science (Wash DC) 215:677–679
Graham F, Van der Eb L (1973) A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467
Häuser H, Gross G, Bruns W, Hochkeppel HK, Mayr U, Collins J (1982) Inducibility of human 3-interferon gene in mouse L-cell clones. Nature (Lond) 297:650–655
Hohn B (1979) In vitro packaging of λ and cosmid DNA. In: Wu R (ed) Methods in Enzymology, vol 68. Academic Press, New York, p 299
Jaenichen HR, Pech M, Lindenmaier W, Wildgruber N, Zachau HG (to be published 1984) Human VK gene sequences and their implication for V gene evolution. Nucleic Acids Res
Kavathas P, Herzenberg LA (1983) Stable transformation of mouse L cells for human membrane T-cell differentiation antigens and β2-µicro-globulin: Selection by fluorescence-activated cell sorting. Proc Natl Acad Sci USA 80:524–528
Kurtz DT (1981) Hormonal inducibility of rat β2µ globulin genes in transfected mouse cells. Nature (Lond) 291:629–631
Lai EC, Woo SLC, Bordelon-Riser ME, Fraser TH, O’Malley BA (1980) Ovalbumin is synthesized in mouse cells transformed with the natural chicken ovalbumin gene. Proc. Natl Acad Sci USA 77:244–248
Lindenmaier W, Hauser H, Greiser de Wilke I, Schütz G (1982) Gene shuttling: moving of cloned DNA into and out of eukaryotic cells. Nucleic Acids Res 10:1243–1256
Liu C-P, Tucker PW, Mushinski JF, Blattner FR (1980) Mapping of heavy chain genes for mouse immunoglobulins M and D. Science (Wash DC) 209:1348–1353
Lowy I, Pellicer A, Jackson JF, Sim GK, Silverstein S, Axel R (1980) Isolation of transforming DNA: cloning of the hamster apart gene. Cell 22:817–823
Lund B, Edlund T, Lindenmaier W, Ny T, Collins J, Lundgren E, von Gabain A (1984) Novel cluster of α-interferon gene sequences in a placental cosmid DNA library. Proc Natl Acad Sci USA 81:2435–2439
Lund T, Grosveld FG, Flavell RA (1982) Isolation of transforming DNA by cosmid rescue. Proc Natl Acad Sci USA 79:520–524
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor New York, pp 80–84
Mayo KE, Warren R, Palmiter RD (1982) The mouse metallothionein-I gene is transcriptionally regulated by cadmium following transfection into human or mouse cells. Cell 29:99–108
Perucho M, Hanahan D, Lipsich L, Wigler M (1980a) Isolation of the chicken thymidine kinase gene by plasmid rescue. Nature (Lond) 285:207–211
Perucho M, Hanahan D, Wigler M (1980b) Genetic and physical linkage of exogenous sequences in transformed cells. Cell 22:309–317
Shih C, Weinberg RA (1982) Isolation of a transforming sequence from a human bladder carcinoma cell line. Cell 29:161–169
Wigler M, Silverstein S, Lee L-S, Pellicer A, Cheng Y, Axel R (1977) Transfer of purified herpes simplex virus thymidine kinase gene to cultured mouse cells. Cell 11:223–232
Wigler M, Pellicer A, Silverstein S, Axel R (1978) Biochemical transfer of single-copy eukaryotic genes using total cellular DNA as donor. Cell 14:725–731
Wold B, Wigler M, Lacy E, Maniatis T, Silverstein S, Axel R (1979) Introduction and expression of a rabbit-β-globin gene in mouse fibroblasts. Proc Natl Acad Sci USA 76:5684–5688
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1984 Springer-Verlag Berlin Heidelberg
About this paper
Cite this paper
Lindenmaier, W., Hauser, H., Collins, J. (1984). An Alternative Gene Cloning Method for the Isolation of Human Genes by Expression in Mouse Cell Clones. In: Schell, J.S., Starlinger, P. (eds) The Impact of Gene Transfer Techniques in Eukaryotic Cell Biology. 35. Colloquium der Gesellschaft für Biologische Chemie 12.–14. April 1984 in Mosbach/Baden, vol 35. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-70065-1_2
Download citation
DOI: https://doi.org/10.1007/978-3-642-70065-1_2
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-70067-5
Online ISBN: 978-3-642-70065-1
eBook Packages: Springer Book Archive