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当前位置: 首页 > 文章 > 水貂阿留申病毒部分VP2基因的克隆表达和重组蛋白的免疫学分析 吉林农业大学学报 2009,31 (3) 330-333
Position: Home > Articles > Cloning and Expression of Region VP2 Gene of Aleutian Mink Disease Parvovirus and Analysis of Immunogenicity of the Recombinant Protein Journal of Jilin Agricultural University 2009,31 (3) 330-333

水貂阿留申病毒部分VP2基因的克隆表达和重组蛋白的免疫学分析

作  者：
孟庆峰;梁艳婷;肖成蕊;宋战昀;钱爱东;王伟利
单  位：
吉林出入境检验检疫局;吉林农业大学动物科学技术学院
关键词：
水貂阿留申病毒;VP2基因;克隆表达;重组蛋白
摘  要：
根据GenBank中已发表的水貂阿留申病毒(ADV)VP2基因核苷酸序列设计合成1对特异引物,利用PCR方法扩增ADV国内分离株部分VP2基因片段,将其克隆到原核表达载体pET-28a中。经酶切、PCR扩增和测序分析证实其已正确插入到表达载体中,构建了原核表达载体pET-28a-VP2。阳性重组质粒转化宿主菌BL21,用IPTG进行诱导表达,对表达产物进行SDS-PAGE和免疫印迹分析。结果表明蛋白获得了表达,表达产物的分子质量为21 kD,与理论推测的分子质量一致;在终浓度为1 mmol/L的IPTG诱导下,5 h时表达量达到高峰;Western blot分析表明表达蛋白能被兔抗水貂抗体所识别,具有相应的抗原性。
译  名：
Cloning and Expression of Region VP2 Gene of Aleutian Mink Disease Parvovirus and Analysis of Immunogenicity of the Recombinant Protein
作  者：
MENG Qing-feng1,2,LIANG Yan-ting2,XIAO Cheng-rui1,SONG Zhan-yun1,QIAN Ai-dong2,WANG Wei-li11.Jilin Entry-Exit Inspection and Quarantine Bureau,Changchun 130062,China; 2.College of Animal Science and Technology, Jilin Agricultural University,Changchun 130118,China
关键词：
Aleutian mink disease parvovirus;VP2 genes;cloning and expression;recombinant protein
摘  要：
The primers were designed according to the complete genome sequences of Aleutian mink disease parvovirus(ADV) published in GenBank.The part domains of VP2 gene of ADV strain were amplified by PCR and cloned into pET-28a vector.PCR,restriction endonuclease digestion and sequence analysis proved the insertion position. It was indicated that the prokaryotic expression vectors Pet-28a-VP2 were constructed.The positive recombinant vectors were transformed into recipient germs BL21 for expression by IPTG inducing.The expressed proteins were measured by SDSPAGE and Western blot.Results showed that the proteins could be expressed successfully.The molecular weights of the expressed proteins were separately 21 kD which agreed with the theory presumed.Induced by IPTG at a final concentration of 1 mmol/L,the expressed protein reached the highest quantity after 5 hours of induction.The Western blot results indicated that the expressed antigen proteins could be recognized by the rabbitanti-Mink antiserum and had some immunoreactivity,and could be efficiently expressed in the prokaryotic system with immunological activities.

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