| 猪圆环病毒ZZ株ORF2基因的克隆及原核表达 |
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| 引用本文： | 乔宏兴,张晓根,边传周,宁豫昌.猪圆环病毒ZZ株ORF2基因的克隆及原核表达[J].动物医学进展,2012(1):33-36. |
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| 作者姓名： | 乔宏兴 张晓根 边传周 宁豫昌 |
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| 作者单位： | 郑州牧业工程高等专科学校生物工程系 |
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| 基金项目： | 河南省科技成果转化项目(092201110008) |
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| 摘 要： | 对所分离鉴定的猪圆环病毒2型(PCV-2)ZZ株,参照GenBank中PCV-2全基因序列,设计1对特异性引物,利用PCR扩增出PCV-2ORF2片段,原核表达载体pET-32a(+)获得重组质粒pET-ORF2,IPTG进行诱导表述,并进行Western blot分析。结果表明,PCR扩增产物长度为702bp,大小与预期的一致,成功构建重组质粒,经IPTG诱导成功获得分子质量约为30ku的表达蛋白,该蛋白具有很好的免疫原性,为进一步研制PCV-2疫苗提供依据。
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| 关 键 词： | 猪原环病毒2型 ORF2基因 原核表达 |
Prokaryotic Expression of ZZ strain ORF2 Gene of PCV-2 |
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| QIAO Hong-xing,ZHANG Xiao-gen,BIAN Chuan-zhou,NING Yu-chang.Prokaryotic Expression of ZZ strain ORF2 Gene of PCV-2[J].Progress In Veterinary Medicine,2012(1):33-36. |
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| Authors: | QIAO Hong-xing ZHANG Xiao-gen BIAN Chuan-zhou NING Yu-chang |
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| Institution: | (Biology department of Zhengzhou College of Animal Husbandry Engineering,Zhengzhou,Henan,450011,China) |
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| Abstract: | In this research,zz strain of porcine circovirus type2(PCV-2) was isolated from one sample with clinical signs of PMWS in Henan province.According to the gene sequence of PCV-2 published from GenBank,we designed a pair of primers for cloning ORF2 sequence,then the amplified products were cloned into pET-32a(+)to construct recombinant plasmid pET-ORF2 for expression after induction by IPTG,then expressed protein was analysed by Western blot.The result showed that this gene were 702 bp in length,recombinant plasmid was constructed successfully.A 30 ku fusion protein was expressed by IPTG induction and had a good immunogenicity with PCV-2 by Western blot.This research will make a foundation for PCV-2 vaccine developing. |
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| Keywords: | PCV-2 ORF2 prokaryotic expression |
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