Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Transferase other than ribonuclease
Patent
1985-05-06
1990-07-24
Huleatt, Jayme A.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Transferase other than ribonuclease
435320, 43525233, 935 14, C12N 912, C12N 120, C12N 1500
Patent
active
049435310
ABSTRACT:
This invention provides a plasmic which, when introduced into a suitable host cell and grown under appropriate conditions, effects expression of a gene on the plasmid and production of a polypeptide having reverse transcriptase activity. The plasmid is a double-stranded DNA molecule which includes in a 5' to 3' order the following: a DNA sequence which includes an inducible promoter; a DNA sequence which includes an ATG initiation condon; the central portion of the Moloney murine leukemia virus (MuLV) pol gene, said central portion including a DNA sequence which encodes the polypeptide having reverse transcriptase activity; a DNA sequence which contains a gene associated with a selectable or identifiable phenotypic trait which is manifested when the vector is present in the host cell; and a DNA sequence which contains an origin of replication from a bacterial plasmid capable of autonomous replication in the host cell.
The invention also concerns a method for recovering purified enzymatically-active polypeptide having reverse transcriptase activity, the polypeptide being encoded by the plasmid pB6 B15.23, from a suitable host cell e.g., E. coli HB101 producing the polypeptide. Finally, the invention concerns use of the polypeptide to prepare complementary DNA (cDNA).
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Goff Stephen P.
Roth Monica J.
Tanese Naoko
Huleatt Jayme A.
The Trustees of Columbia University in the City of New York
White John P.
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