| 猪圆环病毒Ⅱ型ORF2部分基因的克隆及原核表达 |
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| 引用本文： | 王伟利,边少国,肖成蕊,孟庆峰,刘和平.猪圆环病毒Ⅱ型ORF2部分基因的克隆及原核表达[J].吉林农业大学学报,2009,31(6). |
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| 作者姓名： | 王伟利 边少国 肖成蕊 孟庆峰 刘和平 |
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| 作者单位： | 1. 吉林出入境检验检疫局,长春,130062
2. 辽宁益康生物制品有限公司,沈阳,111000 |
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| 基金项目： | 国家质检总局项目(2005IK042) |
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| 摘 要： | 根据发表的猪圆环病毒Ⅱ型(PCV2)ORF2基因序列,设计合成1对特异性引物,从分离到的PCV2吉林株(JL01)中扩增出PCV2 ORF2579 bp的核苷酸片段,克隆到表达载体pET-32a中,经BamHⅠ和HindⅢ酶切及序列分析获得阳性重组表达质粒pET-32a-ORF2。将其转化到表达宿主菌BL21中,经IPTG诱导,成功表达了ORF2基因编码的部分结构蛋白,分子量为40 kD。通过SDS-PAGE和Western检测表明,表达的重组蛋白能够被PCV2阳性血清所识别,具有良好的反应原性。
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| 关 键 词： | 猪圆环病毒Ⅱ型 ORF2 克隆 原核表达 |
Cloning and Prokaryotic Expression of ORF2 Gene of Porcine Circovirus Type 2 |
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| WANG Wei-li,BIAN Shao-guo,XIAO Cheng-rui,MENG Qing-feng,LIU He-ping.Cloning and Prokaryotic Expression of ORF2 Gene of Porcine Circovirus Type 2[J].Journal of Jilin Agricultural University,2009,31(6). |
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| Authors: | WANG Wei-li BIAN Shao-guo XIAO Cheng-rui MENG Qing-feng LIU He-ping |
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| Abstract: | Based on the published nucleotide sequence of ORF2 gene of porcine circovirus 2, a pair of primers were designed and synthesized. The 579 bp was amplified by polymerase chain reaction(PCR) from Jilin strain, and then the fragment was cloned into pET-32a vector, BamHI and Hind HI restriction endonuclease analysis and DNA sequencing were used to identify the recombinant plasmid of pET-32a-ORF2. A recombinant vector was transformed into E. Coli BL21 cells. Then about 40kD fusion protein was expressed in recombinant strain BL21 after being induced by IPTG.The expressed protein confirmed by SDS - PAGE and Western blotting analysis of reactionogenicity can react with the polyclonal antibody against PCV2. |
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| Keywords: | porcine circovirus type 2 ORF2 gene cloning prokaryotic expression |
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