2型登革病毒ns1基因克隆及其表达产物的生物学特性研究  

Cloning and Expression of the Ns1 Gene Encoding the Nonstructural Protein of Dengue Virus Serotype 2 and Identification of Their Immunogenicity

作　　者：但妍[1] 张娟[1] 张君[1] 郑建[1] 黄爱龙[1] 

机构地区：[1]重庆医科大学附属第二医院病毒性肝炎研究所,感染性疾病教育部重点实验室,重庆400016

出　　处：《生物技术通报》2008年第2期168-171,共4页Biotechnology Bulletin

摘　　要：目的克隆并表达2型登革病毒非结构蛋白ns1基因片段,初步鉴定重组蛋白的生物学特性。方法利用登革热2型病毒重组质粒,经PCR方法扩增出ns1全长基因片段,在pQE30表达系统中表达,表达产物用Ni柱亲和层析纯化后,用鼠抗登革病毒免疫血清对重组蛋白进行Western Blot及ELISA鉴定。结果构建的重组质粒pQE-30/NS1,pQE-30/NS1-N,pQE-30/NS1-80-200aa和pQE-30/NS1-C经IPTG诱导,重组蛋白高效表达并纯化成功,经Western Blot及ELISA证实重组蛋白可以被免疫血清特异识别。结论2型登革病毒结构蛋白表达载体在大肠杆菌SG13009中高效表达。纯化产物具有较强的免疫原性,为进一步研究NS1的生物学特性和血清学检测奠定了基础。Objection To clone and express the nonstructural protein encoded by nsl gene of dengue vires serotype 2 （DEN2）and to identify the immunogenicity of the recombinant proteins, Method The nsl cDNA fragments were amplified with PCR from the plasmid contained the genomic-length cDNA of DEN2 strain New Guinea C （NGC）,and were inserted to the multi-cloning sites of plasmid pQE30 vector and expressed with induction of IPTG, After purification used Ni-NTA,the innnunogenicity of the recombinant proteins identified with rat antiserum against dengue virus by Western blot assay. Result The recombinant proteins were highly expressed in E.coli SG13009 and could be recognized by the sera against dengue virus from rat. Conclusion It is evident that the recombinant proteins show good immunogenicity ,which may provide for a potential source to study the biological significance of the proteins.

关 键 词：登革病毒 非结构蛋白 克隆 原核表达 免疫原性 

分 类 号：Q939.4[生物学—微生物学]