Literature DB >> 2412939

Cloning and overexpression of Moloney murine leukemia virus reverse transcriptase in Escherichia coli.

M L Kotewicz, J M D'Alessio, K M Driftmier, K P Blodgett, G F Gerard.   

Abstract

A pBR322-derived expression vector, plasmid pKD1, was constructed containing the strong leftward promoter (pL) of bacteriophage lambda, the ribosome-binding site (RBS) of the cII gene of lambda, and a unique downstream NdeI restriction site for construction of an ATG initiation codon. The section of the pol gene of Moloney murine leukemia virus (M-MLV) that codes for reverse transcriptase (RT) was cloned into the NdeI site of this vector generating the plasmid pRT103. Upon thermal induction, enzymatically active RT was expressed in Escherichia coli [pRT103]. The identity of this activity was confirmed by its template specificity and its sensitivity to inhibition by immunoglobulin G (IgG) prepared against authentic murine RT. RT represented 20% of the newly synthesized protein in these cells 20 min after induction.

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Year:  1985        PMID: 2412939     DOI: 10.1016/0378-1119(85)90003-4

Source DB:  PubMed          Journal:  Gene        ISSN: 0378-1119            Impact factor:   3.688


  19 in total

1.  Reverse transcriptase. The use of cloned Moloney murine leukemia virus reverse transcriptase to synthesize DNA from RNA.

Authors:  G F Gerard; D K Fox; M Nathan; J M D'Alessio
Journal:  Mol Biotechnol       Date:  1997-08       Impact factor: 2.695

2.  The role of Moloney murine leukemia virus RNase H activity in the formation of plus-strand primers.

Authors:  A J Rattray; J J Champoux
Journal:  J Virol       Date:  1987-09       Impact factor: 5.103

3.  Expression of soluble, enzymatically active, human immunodeficiency virus reverse transcriptase in Escherichia coli and analysis of mutants.

Authors:  A Hizi; C McGill; S H Hughes
Journal:  Proc Natl Acad Sci U S A       Date:  1988-02       Impact factor: 11.205

4.  Isolation of cloned Moloney murine leukemia virus reverse transcriptase lacking ribonuclease H activity.

Authors:  M L Kotewicz; C M Sampson; J M D'Alessio; G F Gerard
Journal:  Nucleic Acids Res       Date:  1988-01-11       Impact factor: 16.971

5.  Functional organization of the murine leukemia virus reverse transcriptase: characterization of a bacterially expressed AKR DNA polymerase deficient in RNase H activity.

Authors:  J G Levin; R J Crouch; K Post; S C Hu; D McKelvin; M Zweig; D L Court; B I Gerwin
Journal:  J Virol       Date:  1988-11       Impact factor: 5.103

6.  The alpha and beta chains of avian retrovirus reverse transcriptase independently expressed in Escherichia coli: characterization of enzymatic activities.

Authors:  D A Soltis; A M Skalka
Journal:  Proc Natl Acad Sci U S A       Date:  1988-05       Impact factor: 11.205

Review 7.  The purification of eukaryotic polypeptides synthesized in Escherichia coli.

Authors:  F A Marston
Journal:  Biochem J       Date:  1986-11-15       Impact factor: 3.857

8.  The role of template-primer in protection of reverse transcriptase from thermal inactivation.

Authors:  Gary F Gerard; R Jason Potter; Michael D Smith; Kim Rosenthal; Gulshan Dhariwal; Jun Lee; Deb K Chatterjee
Journal:  Nucleic Acids Res       Date:  2002-07-15       Impact factor: 16.971

9.  Sulphydryl groups in the template-primer-binding domain of murine leukaemia virus reverse transcriptase. Identification and functional analysis of cysteine-90.

Authors:  S Basu; A Basu; M J Modak
Journal:  Biochem J       Date:  1993-12-15       Impact factor: 3.857

10.  Sensitive oligonucleotide ligation assay for low-level detection of nevirapine resistance mutations in human immunodeficiency virus type 1 quasispecies.

Authors:  Matthew S Lalonde; Ryan M Troyer; Aslam R Syed; Stanley Bulime; Korey Demers; Francis Bajunirwe; Eric J Arts
Journal:  J Clin Microbiol       Date:  2007-06-13       Impact factor: 5.948

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