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| 牛疙瘩皮肤病病毒ORF132基因的克隆及其原核表达 | |
| 引用本文： | 周雪梅,聂福平,王昱,肖进文,聂奎,李应国.牛疙瘩皮肤病病毒ORF132基因的克隆及其原核表达[J].中国预防兽医学报,2012,34(4):320-322. |
| 作者姓名： | 周雪梅 聂福平 王昱 肖进文 聂奎 李应国 |
| 作者单位： | 1. 西南大学动物科技学院,重庆北碚400715;重庆出入境检验检疫局,重庆江北400020 2. 重庆出入境检验检疫局,重庆江北400020;重庆进出口食品安全工程中心,重庆江北400020 3. 西南大学动物科技学院,重庆北碚,400715 |
| 基金项目： | 国家质检总局科研项目(2010IK0015);国家质检总局科研项目(2009IK027) |
| 摘 要： | 为原核表达牛疙瘩皮肤病病毒(LDSV) ORF132并对该基因进行生物信息学分析,本研究采用PCR扩增LSDV的ORF132基因,将其亚克隆至pGEX-6p-1载体中,构建重组质粒pGEX-LSDV-ORF132.生物信息学分析表明,LSDV的ORF132与Neethling vaccine LW1959株LW132、NI-2490株LSDV132和NW-LW株LD132的相应推导氨基酸同源性分别为100％、100％和94.4％,与山羊痘病毒、绵羊痘病毒相应推导氨基酸的同源性为20.7 ％～36％.SDS-PAGE和western blot分析表明,pGEX-LSDV-ORF 132在大肠杆菌BL21中经IPTG诱导主要以包涵体形式存在,其重组蛋白大小约为48.95ku,并且与特异性抗体具有抗原反应活性. |
| 关 键 词： | 牛疙瘩皮肤病病毒 LSDV-ORF132基因 序列分析 原核表达 |
Cloning and prokaryotic expression of ORF132 of Lumpy skin disease virus |
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| ZHOU Xue-mei , NIE Fu-ping , WANG Yu , XIAO Jin-wen , NIE Kui , LI Ying-guo.Cloning and prokaryotic expression of ORF132 of Lumpy skin disease virus[J].Chinese Journal of Preventive Veterinary Medicine,2012,34(4):320-322. | |
| Authors: | ZHOU Xue-mei NIE Fu-ping WANG Yu XIAO Jin-wen NIE Kui LI Ying-guo |
| Institution: | 2,3(1.College of Animal Science and Technology,Southwest China University,Chongqing 400715,China; 2.Chongqing Entry-Exit Inspection and Quarantine Bureau of China,Chongqing 400020,China; 3.Chongqing Import and Export Food Safety Engineering Center,Chongqing 400020,China) |
| Abstract: | In response to the risk of Lumpy skin disease virus(LSDV) spreading to China,it is necessary to establish inspection methods for LSDV infection.Through bioinformatics analysis,it was suggested that LSDV-ORF132 shared high homology with LW132(Neethling vaccine),LSDV132(NI-2490) and LW132(NW-LW),the amino acid homologies were 100%,100% and 94%,respectively.In comparison with corresponding amino acid sequences of the original gene in capripox virus,the homologies were ranged from 20.7% to 36%.In this experiment,a 542 bp fragment of LSDV-ORF132 was amplified by PCR and inserted into prokaryotic expression vector pGEX-6p-1 for expressed in E.coli.The recombinant protein was 48.95 ku,and specifically reacted with LSDV antibody,indicating the recombinant protein possessed antigenic activity. |
| Keywords: | Lumpy skin disease virus LSDV-ORF132 gene sequence analysis prokaryotic expression |
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