A two-plasmid system for transient expression of cDNAs in primate cells.
This two-plasmid system for transient gene expression can be used in a wide variety of primate cells. It consists of a cDNA expression vector and a helper plasmid. The cDNA cloned in the expression vector is transcribed by the powerful major immediate-early promoter of murine cytomegalovirus. A segment of the rabbit beta-globin gene placed downstream of the cDNA provides signals for splicing and polyadenylation. The helper plasmid provides SV40 T antigen and adenovirus VA RNA. The T antigen induces replication of the expression vector, which contains an SV40 origin of replication, and VA RNA enhances translation of the transcribed mRNA. In monkey kidney cells, with human tissue-type plasminogen activator (t-PA) cDNA as reporter gene, the helper plasmid boosted t-PA production 30-fold and up to 500 ng/ml t-PA accumulated in the medium in the 5 days following transfection of the two plasmids. In nonprimate cells the helper plasmid stimulated expression 3- to 5-fold.
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Biotechnology Department, Ciba-Geigy Ltd., Basel, Switzerland.
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D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D003587:Cytomegalovirus / Q000235:genetics; D004247:DNA / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D007668:Kidney / Q000166:cytology; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection
D000952:Antigens, Polyomavirus Transforming; D004247:DNA