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| Cloning and Expression of argR gene from Corynebacterium crenatum and Arginine Production with the Recombinant Strain |
| JIAO Hai-tao1, YUAN Yong2, XU Feng2, YANG Wei1, XIONG Yong-hua2, CHEN Xue-lan1 |
1. College of Life Science, JiangXi Normal University, Nanchang 330022, China;
2. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China |
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Abstract Corynebacterium crenatum AS. M7, mutated and stored is a high arginine producing strain. ArgR is one of the most important regulating proteins during arginine biosynthesis. To further confirm the effect of ArgR on arginine biosynthesis in C. crenatum, specific primers were designed and used for PCR amplification of full argR gene from wild strain C. crenatum AS 1.542 and a mutant strain C. creantum AS. M7. The amplified argR gene were aligned and compared for difference. Result showed that argR from C. crenatum AS 1.542 has ORF of 516 bp, encoding a protein consists of 172 amino acids. In comparison, argR from C. crenatum AS. M7 has a C instead of T on position 109, resulting termination of ArgR expression in C. crenatum AS. M7. Meanwhile, argR gene from C. crenatum AS 1.542 was cloned into pXMJ19 vector and transformed into C. crenatum AS. M7 to form a recombinant strain. Arginine production with recombinant strain was evaluated by fermentation in triangle flask. SDS-PAGE and Western blot were used for argR gene expression analysis, result showed a decrease of arginine production from 7.8 mg/ml to 2.5 mg/ml (appr. 67.9%).
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Received: 29 June 2011
Published: 25 October 2011
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