Transformation-associated recombination cloning

US6391642B1 · kind B1 · utility

Assignee

• The United States of America, as represented by the Department of Health and Human Services · US

Inventors

• Michael A. Resnick · Chapel Hill, US
• Vladimir L. Larionov · Potomac, US
• Natalay Kouprina · Potomac, US
• Edward Perkins · Savannah, US

Key dates

• Filing date: Apr 14, 1998
• Grant date: May 21, 2002
• Expiry date: Apr 14, 2018

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N15/81
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The invention is directed to a method of making a yeast artificial chromosome (YAC) comprising introducing into yeast cells a population of nucleic acids and a vector, wherein the vector comprises a yeast centromere, a selectable marker, a yeast telomere, and a sequence which can recombine with a region of a nucleic acid within the population of nucleic acids, whereby in vivo recombination makes the YAC. The invention is also directed to a method of making a YAC using two vectors and a method of making a circular YAC. The invention is also directed toward methods of making YACs with a selected nucleic acid insert from a mixed population of nucleic acids using transformation-associated recombination. The invention is further directed toward a method of cloning a selected nucleic acid from a population of nucleic acids into a vector comprising introducing into yeast cells a population of nucleic acids and the vector, wherein the vector comprises a specific sequence which can recombine with a region of the selected nucleic acid within the population of nucleic acids and a non-specific sequence which can recombine with the selected nucleic acid within the population of nucleic acids; w…