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Science 26 March 1993:
Vol. 259. no. 5103, pp. 1904 - 1907
DOI: 10.1126/science.8096090
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Articles

Science, Vol 259, Issue 5103, 1904-1907
Copyright © 1993 by American Association for the Advancement of Science

articles

Germ line transmission of a yeast artificial chromosome spanning the murine alpha 1(I) collagen locus

WM Strauss, J Dausman, C Beard, C Johnson, JB Lawrence, and R Jaenisch

Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge 02142.

Molecular complementation of mutant phenotypes by transgenic technology is a potentially important tool for gene identification. A technology was developed that allows the transfer of a physically intact yeast artificial chromosome (YAC) into the germ line of the mouse. A purified 150-kilobase YAC encompassing the murine gene Col1a1 was efficiently introduced into embryonic stem (ES) cells via lipofection. Chimeric founder mice were derived from two transfected ES cell clones. These chimeras transmitted the full length transgene through the germ line, generating two transgenic mouse strains. Transgene expression was visualized as nascent transcripts in interphase nuclei and quantitated by ribonuclease protection analysis. Both assays indicated that the transgene was expressed at levels comparable to the endogenous collagen gene.


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