猪去乙酰化酶SIRT3的克隆及表达

Cloning and Expression of Sus scrofa Sirtuin 3

WANG Hou-guang¹, MA Miao-peng², HUANG Kui-ying³, LI Hua-zhou¹, MING Fei-ping³, WANG Wei-fang¹, XIA Feng-geng³, LUO Meng-xiao², YANG Jun², CAI Hai-ming², SHI Ju-qing², HUANG Chao-yuan², CHU Pin-pin², DONG Jian-ming¹,ZHU Hong-xia¹,ZHANG Ling-hua²

(1. Guangzhou Fine Breed Swine Farm, Guangzhou 510540, China; 2. Key Laboratory of Function and Regulation of Agricultural Biotechnology Protein of Guangdong Province, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China; 3. Guangzhou Institute of Microbiology, Guangzhou 510663, China)

Revised:2013-12-20 Online:2014-05-20 Published:2014-06-25

Abstract

Abstract: The study was aimed to obtain recombinant Sus scrofa sirtuin 3 (SIRT3) in prokaryotic system and preliminarily evaluate its antigenicity and specificity. The fragment which included the SIRT3 complete coding sequence was obtained by RT-PCR and cloned into pET-28a(+) vector, then the recombinant vector was transformed into E.coil Rosetta （DE3） to induce, express and purify, the antigenicity of recombinant SIRT3 was identified by Western blotting. The double enzyme analysis and sequencing proved that recombinant plasmid pET28a-SIRT3 was constructed correctly. The expressed recombinant protein, with a relative molecular mass of about 39 ku, was recognized by positive serum antibody. The result showed that the SIRT3 was successfully expressed in E.coil, which would provide a useful tool for designing an in-depth investigation of the role of SIRT3.

Key words:

Sus scrofa; SIRT3; prokaryotic expression

WANG Hou-guang,MA Miao-peng,HUANG Kui-ying,LI Hua-zhou, ING Fei-ping,WANG Wei-fang,XIA Feng-geng,LUO Meng-xiao, ANG Jun,CAI Hai-ming,SHI Ju-qing,HUANG Chao-yuan,CHU Pin-pin,DONG Jian-ming,ZHU Hong-xia. Cloning and Expression of Sus scrofa Sirtuin 3[J]. .

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Cloning and Expression of Sus scrofa Sirtuin 3

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