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Nucleic Acids Research, 1991, Vol. 19, No. 2 319-325
© 1991

MOLECULAR BIOLOGY

The use of engineered E1A genes to transactivate the hCMV-MIE promoter in permanent CHO cell lines

Mark I. Cockett*, Christopher R. Bebbington and Geoffrey T. Yarranton

Celltech Ltd 216 Bath Road, Slough, SL1 4EN, UK

*To whom correspondence should be addressed

Received October 9, 1990. Revised December 10, 1990. Accepted December 10, 1990.

Vectors expressing adenovirus 5 E1A or a domain 2 mutant E1A were introduced into CHO-K1 cells in order to transactivate the hCMV-MIE promoter in transient and stable transfections. Expression from the hCMV promoter was efficiently activated by both wild-type and mutant E1A in contrast to other viral promoters such as the SV40 early promoter which are repressed by E1 A. E1A genes expressed from a strong promoter were inhibitory to the growth of CHO cells. Nevertheless, by the use of a weaker promoter, it was possible to Isolate stably transfected cell lines containing a level of E1A compatible with both continued cell growth and significant transactlvation of the hCMV promoter. By this means we have generated cell lines secreting tissue Inhibitor of metallo-proteinases (TIMP) at levels approaching those previously attained using gene amplification. CHO cell lines constitutively expressing wild-type and mutant E1A genes have been derived which can serve as new host cell lines for transient expression and efficient stable expression without gene amplification.


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