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

P Gunning; J Leavitt; G Muscat; S Y Ng; L Kedes

doi:10.1073/pnas.84.14.4831

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

P Gunning·J Leavitt·G Muscat·S Y Ng·L Kedes

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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.

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Actins Cell Line Fibroblasts Gene Expression Regulation Genetic Vectors Humans Plasmids Promoter Regions, Genetic RNA RNA, Antisense Recombinant Fusion Proteins Simian virus 40 Tubulin Comparative Study Journal Article 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.

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P Gunning, J Leavitt, G Muscat, S Y Ng, L Kedes. 1987. A human beta-actin expression vector system directs high-level accumulation of antisense transcripts.. https://doi.org/10.1073/pnas.84.14.4831

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