羊传染性脓疱病毒趋化因子结合蛋白(CBP)基因的克隆及表达分析  

Cloning and expression analysis of the chemokine-binding protein(CBP)gene of Orf virus

作　　者：王改丽[1] 赵魁[1,2] 兰云刚[1] 贺文琦[1] 陆慧君[2] 解胜男[1] 苏高莉[1] 张冰冰[1] 宋德光[1] 高丰[1,2] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]吉林大学人兽共患病研究所,吉林长春130062

出　　处：《中国兽医学报》2011年第8期1114-1117,共4页Chinese Journal of Veterinary Science

基　　金：吉林省科技厅重点计划发展项目(20080217);吉林大学基本科研业务费平台基地建设项目(450060326051)

摘　　要：参照GenBank中ORFV的毒力因子CBP基因的核苷酸序列设计合成1对特异性引物,采用PCR方法扩增该基因片段,并克隆入原核表达载体pGEX-4T-1中,经PCR、酶切鉴定和测序正确后,转入E.coli BL21(DE3)感受态细胞中,经IPTG进行诱导表达,并通过生物信息学软件进行分析。SDS-PAGE结果可见大小约为49 000的融合蛋白条带,抗原性分析表明其具有良好的免疫原性;此外,Western-blotting试验结果确证了GST-CBP融合蛋白的表达并证实其具有良好的免疫原性。结果表明,本研究成功诱导表达GST-CBP融合蛋白,这将为进一步研究CBP的功能及理化特性,以及开发以CBP为基础的诊断抗原奠定了物质基础。According to the published sequence of Orf virus（ORFV） virulence factors CBP genein Genbank,a pair of primers was designed and synthesized.The sequence of CBP gene was amplified by PCR.The amplicon was inserted into pGEX-4T-1 vector,and the recombinant plasmid was identified by PCR and digestion with restriction endonucleases,and then sequenced.The positive recombinant plasmid was transformed into E.coli BL21（DE3） and was induced by IPTG.Then,the expressed protein was analysed through bioinformatics software.The result of SDS-PAGE electrophoresis showed that expressed fusion protein was about 49 KDa.And the fusion protein had favourable antigenicity by antigenic analysis.In addition,the GST-CBP fusion protein had the better immunogenicity further confirmed by Western-blotting.The results mentioned above showed that the GST-CBP fusion protein was successfully induced.It would provide the material basis for further study of the function and physical and chemical properties of CBP,and the development of diagnostic antigen of CBP-based.

关 键 词：羊传染性脓疱病毒 趋化因子结合蛋白 克隆 原核表达 

分 类 号：S852.659.1[农业科学—基础兽医学]