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PMID: 2440031 Published · ppublish English 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.

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

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 84 ·No. 14 ·1987-07-00 ·Pages 4831-5

Gunning P, Leavitt J, Muscat G, Ng SY, Kedes L

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.

MeSH Terms
Actins/biosynthesis,genetics Cell Line Fibroblasts/metabolism Gene Expression Regulation Genetic Vectors Humans Plasmids Promoter Regions, Genetic RNA/biosynthesis,genetics RNA, Antisense Recombinant Fusion Proteins/biosynthesis,genetics Simian virus 40/genetics Tubulin/biosynthesis,genetics
Chemicals
Actins RNA, Antisense Recombinant Fusion Proteins Tubulin RNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gunning P
Leavitt J
Muscat G
Ng S Y
Kedes L
References (33)
33 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.84.14.4831
Published
1987-07-00
Pages
4831-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC305199
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.84.14.4831 PMC Full Text View journal details
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