Amplifiable expression vectors for mammalian cell lines.
We have constructed a prototype of a general expression vector for mammalian cells, which carriers a strong promoter active in a variety of tissues and animal species for expression of the gene of interest and a truncated gene for amplification in selective medium. Expression of the S gene of HBV encoding the surface antigen was used to evaluate various versions of this vector. The human metallothionein IIA promoter was found to be particularly efficient for expression of this gene, both in mouse L and monkey Vero cells. Including a promoterless tk gene in the vector led to gene amplification in Ltk- cells by selection of TK+ variants in selective medium containing hypoxanthine, aminopterine and thymidine (HAT medium). Concomitant increases of the S gene expression levels were initially 50-100 fold. Although many clones were unstable, even in selective medium, some maintained the high expression levels for over one year.
D000818:Animals; D002460:Cell Line; D005784:Gene Amplification; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006515:Hepatitis B virus / Q000235:genetics; D008668:Metallothionein / Q000235:genetics; D051379:Mice; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014170:Transformation, Genetic
D008668:Metallothionein; D013937:Thymidine Kinase