| 猪瘟病毒山东惠民分离株E2基因的克隆及表达 |
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| 引用本文： | 魏凤,张文通,王金良,杨慧,沈志强.猪瘟病毒山东惠民分离株E2基因的克隆及表达[J].兽医大学学报,2012(8):1098-1100. |
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| 作者姓名： | 魏凤 张文通 王金良 杨慧 沈志强 |
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| 作者单位： | [1]山东省滨州畜牧兽医研究院,山东滨州256600 [2]山东绿都生物科技有限公司,山东滨州256600 |
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| 基金项目： | 山东省自然科学基金资助项目（ZR2010CQ012） |
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| 摘 要： | 从山东惠民某猪场病料中经RT-PCR扩增了猪瘟病毒（CSFV）E2基因,并将其克隆到pMD18-T载体。经序列测定,E2基因核苷酸序列长度为1 065bp,与GenBank中CSFV石门毒株（SHIMEN）、猪瘟兔化弱毒疫苗株（HCLV）、猪瘟ZJ7分离株E2基因核苷酸序列同源性分别为85.1%,84.2%,98.8%;氨基酸序列同源性分别为91.5%,91.5%,98.6%。将E2基因插入到大肠杆菌原核表达载体pGEX-KG,获得重组质粒pKG-E2,再转化大肠杆菌BL21（DE3）感受态细胞,经IPTG诱导,受体菌能表达大小约为65 000的蛋白。Western blot显示表达的蛋白具有反应原性。
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| 关 键 词： | 猪瘟病毒 E2基因 克隆表达 |
Cloning and expression of E2 gene of classical swine fever virus isolates from Huimin county of Shandong province |
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| Authors: | WEI Feng ＇ ZHANG Wen-tong WANG Jin-liang ＇ YANG Hui SHEN Zhi-qiang |
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| Institution: | 1. Shan dong na ;2. Binzhou Animal Science＆ Veterinary Medicine Academy, Binzhou , Shandong 256600, China;2. Shandong Liidu Biological Technology Co. , Ltd. , Binzhou, Shandong 256600, China) |
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| Abstract: | The structural protein gene E2 was amplified from the genome of classical swine fever virus（CSFV） from a pig farm of Huimin county in Shandong by RT-PCR,and then cloned into pMD18-T vector for sequence analysis. The results of sequence analysis showed that E2 gene has 1 065 base pair. The E2 gene nucleotide sequence and deduced amino acid sequence shared homology with CSFV SHIMIN, HCLV, ZJ7 as follow： the nucleotide sequence 85.1%, 84.2 %, 98.8 % ; and the amino acid sequence 91.5 %, 91.5 %, 98.6 %, respectively. The E2 gene was sub- cloned into the expression vector pGEX-KG to construct pKG-E2, and then transformed into E. coli BL21 （DE3）. E2 protein expression was induced by IPTG. The results of SDS-PAGE indicated that the molecular weight of E2 fusion protein was about 65 000. The results of Western blot showed a reacitivity with the expressed protein. |
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| Keywords: | classical swine fever virus E2 gene cloning and expression |
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