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PMID: 2410910 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Expression of enzymatically active reverse transcriptase in Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 82 ·No. 15 ·1985-08-00 ·Pages 4944-8

Tanese N, Roth M, Goff SP

Abstract

Reverse transcriptase of murine retroviruses is a monomeric protein of approximately 80,000 daltons, which is encoded by the central portion of the viral pol gene. To prepare large quantities of the enzyme, we have constructed gene fusions between the trpE gene and portions of the pol gene of Moloney murine leukemia virus. The inserted pol gene sequences include the entire coding region for the mature enzyme and various amounts of additional coding sequences. Many of these constructs express high levels of reverse transcriptase activity even though the NH2 and COOH termini of the protein product only approximate the correct termini of the authentic protein.

MeSH Terms
DNA, Recombinant Escherichia coli/genetics Gene Expression Regulation Genes, Viral Molecular Weight Moloney murine leukemia virus/enzymology Plasmids RNA-Directed DNA Polymerase/genetics Tryptophan/genetics
Chemicals
DNA, Recombinant Tryptophan RNA-Directed DNA Polymerase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tanese N
Roth M
Goff S P
References (21)
21 references, click to expand
  1. RNA-directed DNA polymerase--properties and functions in oncogenic RNA viruses and cells.
    Prog Nucleic Acid Res Mol Biol. 1974;14(0):187-334 PMID: 4135976
  2. Reverse transcription by Escherichia coli DNA polymerase I.
    Proc Natl Acad Sci U S A. 1973 Dec;70(12):3834-8 PMID: 4129927
  3. The reverse transcriptase.
    Biochim Biophys Acta. 1977 Mar 21;473(1):1-38 PMID: 66065
  4. Further characterization of intracellular precursor polyproteins of Rauscher leukemia virus.
    Virology. 1977 May 1;78(1):11-34 PMID: 67705
  5. Preparative and analytical purification of DNA from agarose.
    Proc Natl Acad Sci U S A. 1979 Feb;76(2):615-9 PMID: 284385
  6. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  7. Isolation and properties of Moloney murine leukemia virus mutants: use of a rapid assay for release of virion reverse transcriptase.
    J Virol. 1981 Apr;38(1):239-48 PMID: 6165830
  8. Cloned viral protein vaccine for foot-and-mouth disease: responses in cattle and swine.
    Science. 1981 Dec 4;214(4525):1125-9 PMID: 6272395
  9. Deletion mutants of Moloney murine leukemia virus which lack glycosylated gag protein are replication competent.
    J Virol. 1983 May;46(2):538-46 PMID: 6601726
  10. Construction of mutants of Moloney murine leukemia virus by suppressor-linker insertional mutagenesis: positions of viable insertion mutations.
    Proc Natl Acad Sci U S A. 1984 Jul;81(13):4149-53 PMID: 6330745
  11. Construction and analysis of deletion mutations in the pol gene of Moloney murine leukemia virus: a new viral function required for productive infection.
    Cell. 1984 Jul;37(3):1043-52 PMID: 6204767
  12. A mutant murine leukemia virus with a single missense codon in pol is defective in a function affecting integration.
    Proc Natl Acad Sci U S A. 1984 Oct;81(20):6461-5 PMID: 6208550
  13. A deletion mutation in the 5' part of the pol gene of Moloney murine leukemia virus blocks proteolytic processing of the gag and pol polyproteins.
    J Virol. 1985 Mar;53(3):899-907 PMID: 3882995
  14. ENZYMATIC SYNTHESIS OF DEOXYRIBONUCLEIC ACID. XIV. FURTHER PURIFICATION AND PROPERTIES OF DEOXYRIBONUCLEIC ACID POLYMERASE OF ESCHERICHIA COLI.
    J Biol Chem. 1964 Jan;239:222-32 PMID: 14114848
  15. A complementation analysis of the restriction and modification of DNA in Escherichia coli.
    J Mol Biol. 1969 May 14;41(3):459-72 PMID: 4896022
  16. RNA-dependent DNA polymerase in virions of RNA tumour viruses.
    Nature. 1970 Jun 27;226(5252):1209-11 PMID: 4316300
  17. RNA-dependent DNA polymerase in virions of Rous sarcoma virus.
    Nature. 1970 Jun 27;226(5252):1211-3 PMID: 4316301
  18. Analysis of DNA polymerases II and 3 in mutants of Escherichia coli thermosensitive for DNA synthesis.
    Proc Natl Acad Sci U S A. 1971 Dec;68(12):3150-3 PMID: 4943556
  19. Isolation and partial characterization of a mutant of Escherichia coli deficient in DNA polymerase II.
    Proc Natl Acad Sci U S A. 1972 Aug;69(8):2090-4 PMID: 4559593
  20. Association of an endoribonuclease with the avian myeloblastosis virus deoxyribonucleic acid polymerase.
    J Biol Chem. 1972 Nov 25;247(22):7282-7 PMID: 4344646
  21. On the origin of RNA tumor viruses.
    Annu Rev Genet. 1974;8:155-77 PMID: 4140706
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.82.15.4944
Published
1985-08-00
Pages
4944-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC390474
Subset
IM
Grants
NCI NIH HHS · 1 R01 CA 30488 · United States
External Links
PubMed source DOI: 10.1073/pnas.82.15.4944 PMC Full Text View journal details
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