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Nucleic Acids Research, 1982, Vol. 10, No. 21 6715-6732
© 1982

MOLECULAR BIOLOGY

The construction of cosmid libraries which can be used to transform eukaryotic cells

F.G. Grosveld, T. Lund, E.J. Murray, A.L Mellor, H.H.M. Dahl and R.A. Flavell

Laboratory of Gene Structure and Expression, National Institute for Medical Research Mill Hill, London NW7 1AA, UK

Received August 10, 1982. Accepted September 21, 1982.

Cosmid vectors have been developed which carry selective markers for growth in bacteria (ß lactamase gene) and animal cells (the Herpes Simplex virus thymidine kinase gene, the transposon Tn–5 aminoglycosyl 3' phosphotransrefase gene and the E. coli guanine phosphoribosyltransferase gene). The design of the cosmids allows the exchange of the eukaryotic markers in recombinant cosmids. Human and mouse cosmid libraries containing DNA inserts of about 40kb have been generated by an improved method. Several clones from the human ß–globin locus were isolated. These cosmids transform mouse L cells at high efficiency in both circular and linear form. The newly introduced genes are expressed accurately in L cells.


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