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中国生物工程杂志

China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (02): 60-65    DOI:
    
Prokaryotic Expression of Major Epitope Domain of African Swine Fever Virus VP73 Gene
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Abstract  

A full-length VP73 gene sequence was synthesized, the its higher antigen area is predicted by using DNAStar software, and one pair of specific primers for this area are designed by using Primer Premier 5.0. Amplified 243bp fragment of the VP73 gene (vp73l) by PCR was digested and cloned into the prokaryotic expression vector pET-32a. The positive clone of inserting VP73L gene with correct reading frame was confirmed by sequencing and colony PCR. After induction by IPTG, the fusion protein was highly expressed in Escherichia coli BL21 (DE3) in soluble form. The recombinant protein was purified with His-Bind affinity chromatography. Western blotting analysis revealed that the recombinant protein could react with rabbit anti-African swine fever virus VP73 polyclonal antibody. Fusion expression of African swine fever virus VP73L is helpful to to prepare ASFVserological diagnostic reagent in next work.



Key wordsAfrican swine fever virus      VP73gene      Major epitope domain      Expresion     
Received: 21 September 2009      Published: 26 February 2010
Corresponding Authors: Jianhua WANG     E-mail: tengda@mail.caas.net.cn,wangjianhua@mail.caas.net.cn
Cite this article:

LI Qiu-Xia, TENG Da, TONG De-Wen, YANG Ya-Lin, TIAN Zi-Gang, WANG Jian-Hua. Prokaryotic Expression of Major Epitope Domain of African Swine Fever Virus VP73 Gene. China Biotechnology, 2010, 30(02): 60-65.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I02/60

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