A human beta-actin expression vector system directs high-level accumulation of antisense transcripts.
We have constructed a mammalian expression vector consisting of 3 kilobases of the human beta-actin gene 5' flanking sequence plus 5' untranslated region and intervening sequence 1 linked at the 3' splice site to a short DNA polylinker sequence containing unique Sal I, HindIII, and BamHI restriction endonuclease sites followed by a simian virus 40 (SV40) polyadenylylation signal. Two derivatives, containing the selection markers obtained from pSV2gpt or pSV2neo, were also generated. We find that the promoter activity of this vector is a great or greater than that of the SV40 early promoter in a variety of human and rodent cells. The vector was used to generate gamma-actin and beta-tubulin antisense transcripts in human fibroblast cell lines. The antisense transcripts accumulate to levels comparable with that of the highly abundant gamma-actin and beta-tubulin mRNAs.
D000199:Actins / Q000096:biosynthesis / Q000235:genetics*; D002460:Cell Line; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012313:RNA / Q000096:biosynthesis* / Q000235:genetics; D016372:RNA, Antisense; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics; D014404:Tubulin / Q000096:biosynthesis / Q000235:genetics
D000199:Actins; D016372:RNA, Antisense; D011993:Recombinant Fusion Proteins; D014404:Tubulin; D012313:RNA