Expression of enzymatically active reverse transcriptase

US4943531A · kind A · utility

Assignee

• The Trustees of Columbia University in the City of New York · US

Inventors

• Stephen P. Goff · Orefield, US
• Naoko Tanese · New York, US
• Monica Roth · New York, US

Key dates

• Filing date: May 6, 1985
• Grant date: Jul 24, 1990
• Expiry date: May 6, 2005

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2740/13022
• WIPO field: Biotechnology
• WIPO sector: Chemistry

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. Final…