Purification of Chinese Sacbrood Virus (CSBV), Gene Cloning and Prokaryotic Expression of its Structural Protein VP1

文献类型： 外文期刊

第一作者： Wu, Pengjie

作者： Wu, Pengjie;Yu, Huimin;Xu, Jin;Wu, Jiangli;Getachew, Awraris;Tu, Yangyang;Guo, Zhanbao;Jin, Hongyan;Xu, Shufa;Jin, Hongyan

作者机构：

关键词： Apis cerana; Chinese Sacbrood Virus; Virus particle; BJMY-CSBV-VP1; Gene cloning; Sequence analysis; Prokaryotic expression

期刊名称：MOLECULAR BIOTECHNOLOGY （ 影响因子：2.695； 五年影响因子：2.303 ）

ISSN： 1073-6085

年卷期： 2018 年 60 卷 12 期

页码：

收录情况： SCI

摘要： The aim of this study was to purify the Chinese Sacbrood Virus Beijing Miyun (BJMY-CSBV) from infected Apis cerana larvae, clone structural protein gene VP1 (named BJMY-CSBV-VP1), and investigate its biological information. The result indicated that the capsid of CSBV is of spherical shape. Gene clone experiment showed that the BJMY-CSBV-VP1 gene sequence comprised 945bp, encoding 315 amino acids with relative molecular weight of 35.59kDa and isoelectric point 9.38 pI. Phylogenetic analysis of amino acid sequences showed that the BJMY-CSBV-VP1 and LNDD_2015 were grouped together. Protein secondary structure prediction showed that the gene contained two -helices, thirteen beta-folds, six polypeptide binding sites, and no disulfide bridge. Simultaneously, the BJMY-CSBV-VP1 was ligated to the expression vector pET32a(+) and then transformed into the Escherichia coli BL21 (DE3) for prokaryotic expression. The optimal expression experiment revealed that the protein was found in the inclusion body. The recombinant protein was successfully purified by washing buffer combined with supersonic fragmentation. In this study, we obtained the purified BJMY-CSBV particles, cloned BJMY-CSBV-VP1 gene, investigated the detailed information of the gene by analyzing the sequence, and obtained the purified recombinant protein, which could help for further understanding of the function of the structural protein gene VP1.

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