WU多瘤病毒衣壳蛋白原核表达及初步分析  

Prokaryotic expression and pilot application of capsid proteins of WU polyomavirns

作　　者：张银辉[1] 王琼[1] 贾雪[1] 林广裕 黄烈[1] 刘建[1] 陆学东[1] 

机构地区：[1]广东医学院附属深圳福田医院检验医学部,518033 [2]汕头医学院附属第二临床医学院儿科

出　　处：《中华微生物学和免疫学杂志》2011年第6期528-531,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的获得WU多瘤病毒重组衣壳蛋白，分析其抗原性，筛选具有诊断价值的抗原。方法采用PCR技术扩增WU多瘤病毒衣壳蛋白VP1、VP2和VP3的基因片段，并将目的基因插入PGEX-20T表达载体，转化大肠杆菌B121（DE3）后诱导表达。用免疫印迹法对表达蛋白进行鉴定和分析并用临床样本进行初步验证。结果重组质粒在BL21（DE3）菌株中经IPTG诱导、SDS—PAGE和免疫印迹鉴定显示在相对分子质量（Mr）69×10^3、63×10^3和56×10^3处出现清晰条带，重组质粒经双酶切鉴定，表达产物经GST标签鉴定均正确。16份呼吸道分泌物WU多瘤病毒核酸阳性血清标本和70份阴性标本免疫印迹分析表明，重组蛋白具有良好的抗原性。结论成功构建了WU多瘤病毒衣壳蛋白VP1、VP2和VP3的表达载体，表达的重组蛋白具有良好的抗原性，为进一步研究提供了资料。Objective To express the capsid proteins of WU polyomavirus（WUPyV） for research and find antigen for diagnostic value. Methods Coding sequences of capsid proteins of WU polyomavirus by PCR were cloned in prokaryotic expression vector PGEX-20T. Recombinant plasmids were transformed into E. coli BI21 （DE3） and induced by IPTG for proteins expression. Recombinant proteins were identified by Western blot. Results SDS-PAGE proved that recombinant proteins showed three bands with molecular relative mass of 69 ×10^3, 63×10^3 and 56 ×10^3. The recombinant proteins were recognized by anti-GST McAb. The antigenicity was tested by Western blot using 16 WU polyomavirus positive and 70 negative sera. Conclusion Recombinant VP1, VP2 and VP3 expressed in E. coli can combine with WUPyV-Ab and have good antigenicity. They can be used for further research.

关 键 词：WU多瘤病毒 衣壳蛋白 原核表达 

分 类 号：R3[医药卫生—基础医学]