Methods for analyzing the insertion capabilities of modified group II introns

US7592161B2 · kind B2 · utility

Assignees

• The Ohio State University Research Foundation · US
• The University of Texas Board of Regents of the University of Texas System · US

Inventors

• Alan M. Lambowitz · Austin, US
• Huatao Guo · Alhambra, US
• Michael Karberg · Austin, US

Key dates

• Filing date: Oct 22, 2002
• Grant date: Sep 22, 2009
• Expiry date: Nov 25, 2023

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N15/90
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention provides a system and methods for analyzing the function of nucleotide integrases and modified group II introns. The system comprises a donor plasmid comprising a wild-type or modified group II intron, a recipient plasmid comprising a DNA recognition site and a promoterless reporter gene downstream of the DNA target site, and a host cell. The method comprises the steps of transforming a host cell with the donor and recipient plasmids, assaying for expression of the reporter gene, isolating plasmid DNA from the cotransformed cells, and analyzing the plasmid DNA to confirm that the group II intron has been inserted into the target sequence. The present invention also provides a method for simultaneously analyzing the activity of two or more modified nucleotide integrases. The present invention also relates to methods of preparing a library of donor plasmids containing a plurality of diverse modified group II intron DNA sequences.