Method of expression cloning

US5654150A · kind A · utility

Assignees

• President and Fellows of Harvard College · US
• Harvard University, Office of Technology and Trademark Licensing · US

Inventors

• Randall W. King · Newton, US
• Kevin D. Lustig · Cambridge, US
• P. Stukenberg · Brookline, US
• Marc W. Kirschner · Newton, US

Key dates

• Filing date: Jun 7, 1995
• Grant date: Aug 5, 1997
• Expiry date: Jun 7, 2015

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12P21/02
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention provides improvements in the expression cloning method for isolating novel cDNA clones. In particular, in the expression cloning method comprising the steps of preparing a cDNA library from a cell that expresses a desired protein; inserting the cDNA library into an expression vector; inserting the cDNA library into bacterial cells and culturing the bacterial cells to produce individual bacterial colonies; collecting pools of a predetermined number of individual bacterial colonies; isolating the cDNAs contained in the pools; expressing proteins encoded by the cDNAs; and detecting the desired protein, positive clones may be efficiently and cost-effectively obtained by collecting pools of about 100 individual bacterial colonies and expressing proteins encoded by the cDNAs in the pools in vitro.