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Title of Journal: Appl Biochem Biotechnol

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Abbravation: Applied Biochemistry and Biotechnology

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Publisher

Humana Press Inc

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DOI

10.1007/bf00542581

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ISSN

1559-0291

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Gene Cloning Overexpression and Characterization

Authors: Hui Luo Lu Fan Yanhong Chang Jinwei Ma Huimin Yu Zhongyao Shen
Publish Date: 2008/08/05
Volume: 160, Issue: 2, Pages: 393-400
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Abstract

A DNA fragment containing the entire coding sequence of nitrilase gene was amplified from Rhodococcus rhodochrous tg1A6 with high nitrilase activity using PCR and sequenced The open reading frame of the nitrilase gene contains 1101 base pairs which encodes a putative polypeptide of 366 amino acid residues The nitrilase gene was cloned into an expression vector pET28a and expressed in an Escherichia coli strain BL21DE3 The enzymatic activity of nitrilase which converts various nitriles to the corresponding carboxylic acids was detected to reach 245 U/ml at 9 h in the recombinant bacteria

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