Methods of making an RNP particle having nucleotide integrase activity

US6001608A · kind A · utility

Assignee

• The Ohio State Research Foundation · US

Inventors

• Alan M. Lambowitz · Austin, US
• Georg Mohr · Heringen (Werra), DE
• Roland Saldanha · Columbus, US
• Manabu Matsuura · Austin, US
• Clifford J. Beall · Columbus, US
• Jiam Yang · Dallas, US
• Steven Zimmerly · Columbus, US
• Huatao Guo · Alhambra, US

Key dates

• Filing date: May 27, 1998
• Grant date: Dec 14, 1999
• Expiry date: May 27, 2018

Classification

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

Abstract

Methods for preparing nucleotide integrases are provided. The nucleotide integrases are prepared by combining in vitro an excised, group II intron RNA, referred to hereinafter as "exogenous RNA", with a group II intron-encoded protein. The exogenous RNA is prepared by in vitro transcription of a DNA molecule which comprises a group II intron sequence. In one embodiment, the group II intron-encoded protein is made by introducing into a host cell a DNA molecule that comprises at least the open reading frame sequence of a group II intron and then expressing the open reading frame sequence in the host cell. The DNA molecule may comprise the open reading frame sequence operably linked to a promoter, preferably an inducible promoter. Thereafter, the cell is fractionated and the protein is recovered and combined in vitro with the exogenous RNA to provide RNP particles having nucleotide integrase activity. In another embodiment, the DNA molecule comprise a group II intron sequence that encodes both a group II intron RNA as well as a group II intron encoded protein. The DNA molecule is then expressed in the host cell to provide RNP particles that comprise the group II intron-encoded protein…