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J. Biol. Chem., Vol. 282, Issue 26, 18929-18936, June 29, 2007

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Directed Evolution of Ribosomal Protein S1 for Enhanced Translational Efficiency of High GC Rhodopseudomonas palustris DNA in Escherichia coli^*

Jeffrey R. Bernstein, Thomas Bulter, Claire R. Shen, and James C. Liao¹

From the Department of Chemical and Biomolecular Engineering and Biomedical Engineering Interdepartmental Program, UCLA, Los Angeles, California 90095

The expression of foreign DNA in Escherichia coli is important in biotechnological applications. However, the translation of genes from GC-rich organisms is inefficient in E. coli.To overcome this problem, we applied directed evolution to E. coli ribosomal protein S1. Two selected mutants enabled 12- and 8-fold higher expression levels from GC-rich DNA targets. General improvements in translation efficiency over a range of genes from Rhodopseudomonas palustris and E. coli was achieved using an S1 mutant selected against multiple genes from R. palustris. This method opens new opportunities for the expression of GC-rich genes in E. coli.

Received for publication, February 16, 2007 , and in revised form, March 16, 2007.

^* This work was supported by NIGMS Grants 1R01GM077625-01 and DOE DE-FG02-01ER63241 from the National Institutes of Health and UCLA-DOE Institute of Genomics and Proteomics. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Tables 1 and 2.

¹ To whom correspondence should be addressed: 5531 Boelter Hall, 420 Westwood Plaza, Los Angeles, CA 90095. Tel.: 310-825-1656; Fax: 310-206-4107; E-mail: liaoj{at}ucla.edu.

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