Methods for preparing nucleotide integrases

US5804418A · kind A · utility

Assignee

• The Ohio State University Research Foundation · US

Inventors

• Alan M. Lambowitz · Austin, US
• Georg Mohr · Heringen (Werra), DE
• Roland Saldanha · Columbus, US
• Manabu Matsuura · Austin, US

Key dates

• Filing date: Nov 19, 1996
• Grant date: Sep 8, 1998
• Expiry date: Nov 19, 2016

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N9/22
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention provides new, improved, and easily manipulable methods for making nucleotide integrases. In one embodiment, the nucleotide integrase is prepared by introducing a DNA molecule which comprises a group II intron DNA sequence into a host cell. The group II intron DNA sequence is then expressed in the host cell such that RNP particles having nucleotide integrase activity are formed in the cell. Such RNP particles comprise an exiced group II intron RNA encoded by the introduced DNA molecule and a group II intron-encoded protein encoded by the introduced DNA molecule. Thereafter, the nucleotide integrase is isolated from the cell. In another embodiment, the nucleotide integrase is prepared by combining in vitro an excised, group II intron RNA, referred to hereinafter as "exogenous RNA", with a group II intron-encoded protein. In another embodiment, the nucleotide integrase is prepared by combining in vitro an excised, group II intron RNA, referred to hereinafter as "exogenous RNA", with an RNA-protein complex which comprises a group II intron-encoded protein. Preferably, the exogenous RNA is prepared by in vitro transcription of a DNA molecule which comprises the gro…