A human beta-actin expression vector system directs high-level accumulation of antisense transcripts

Proc Natl Acad Sci U S A. 1987 Jul;84(14):4831-5. doi: 10.1073/pnas.84.14.4831.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Actins / biosynthesis
  • Actins / genetics*
  • Cell Line
  • Fibroblasts / metabolism
  • Gene Expression Regulation*
  • Genetic Vectors*
  • Humans
  • Plasmids
  • Promoter Regions, Genetic*
  • RNA / biosynthesis*
  • RNA / genetics
  • RNA, Antisense
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Simian virus 40 / genetics
  • Tubulin / biosynthesis
  • Tubulin / genetics

Substances

  • Actins
  • RNA, Antisense
  • Recombinant Fusion Proteins
  • Tubulin
  • RNA