Highly-purified recombinant reverse transcriptase

Details Highly-purified recombinant reverse transcriptase Highly-purified recombinant reverse transcriptase

1997-01-09

1999-08-10

Hendricks, Keith D.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Transferase other than ribonuclease

536 232, 43525233, C12N 912, C12N 1554, C12N 1570

Patent

active

059358335

ABSTRACT:
A plasmid for expression of Moloney Murine Leukemia Virus-derived reverse transcriptase in E. coli cells deficient in the expression of indiginous RNAse activity, a method for purification of the recombinant enzyme, and a composition comprising a cloned and purified reverse transcriptase opimized for use in cDNA and nucleic acid amplification procedures.

REFERENCES:
patent: 4943531 (1990-07-01), Goff et al.
patent: 5017492 (1991-05-01), Kotewicz et al.
Kelly et al., RNase PH is Essential for tRNA Processing and Viability in RNase-deficient Escherichia coli Cells. J. Biol. Chem. 267(23):16015-16018 (1992).
Durwald, H. and H. Hoffman-Berling, Endonuclease I-deficient and ribonuclease I-deficient Escherichia coli mutants. J. Mol. Biol. 34:331-346 (1968).
Hautala, et al., Increased expression of a euraryotic gene in Escherichia coli through stabalization of its messenger RNA. Proc. Natl. Acad. Sci. USA 76(11):5774-5778 (1979).
Hu, et al., Murine leukemia virus pol gene products: analysis with antisera generated against reverse transcriptase and endonuclease fusion proteins expressed in Escherichia coli. J. Virol. 60(1):267-271 (1986).
Kotewicz, et al., Cloning and overexpression of Moloney murine leukemia virus reverse transriptase in Escherichia coli. 35:249-258 (1985).
Hizi, A. and S. Hughes, Expression in Escherichia coli of a Moloney murine leukemia virus reverse transcriptase whose structure closely resembles the viral enzyme. Gene 66:319-323 (1988).
Telesnitsky, A. and S. Goff, RNase H domain mutations affect the interaction between Moloney murine leukemia virus reverse transcriptase and its primer template. Proc. Natl. Acad. Sci. USA 90:1276-1280 (1993).

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