Hyperexpression of rat spermatidal protein TP2 in Escherichia coli by codon optimization and engineering the vector-encoded 5 ' UTR

Meetei, AR and Rao, MRS (1998) Hyperexpression of rat spermatidal protein TP2 in Escherichia coli by codon optimization and engineering the vector-encoded 5 ' UTR. In: Protein Expression and Purification, 13 (02). pp. 184-190.

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Abstract

We have recently reported the cDNA cloning of rat spermatidal protein TP2 and its expression in Escherichia coli using pTrc 99A as the expression vector. However, the expression level was very low. We have now improved the expression of TP2 over fivefold by (1) optimizing the codons for lysine, arginine, proline, leucine, glycine, valine, threonine, alanine, and tyrosine and (2) by engineering the vector-encoded 5' UTR. The expressed protein was in the soluble phase and could be purified to homogeneity by successive chromatography on Zinc-NTA-agarose affinity matrix and heparin agarose. Serendipitously, we have also observed a concomitant hyperinduction of vector encoded beta-lactamase gene along with TP2 in the E. coli BL21 (DE3) cells.

Item Type:	Journal Article
Publication:	Protein Expression and Purification
Publisher:	Elsevier Science
Additional Information:	Copyright of this article belongs to Elsevier Science.
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	12 Aug 2009 10:37
Last Modified:	19 Sep 2010 05:24
URI:	http://eprints.iisc.ac.in/id/eprint/18698

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