摘要
为获得具有良好抗原性的日本血吸虫重组蛋白,本研究设计了两对特异性引物,以日本血吸虫mRNA为模板,RT-PCR方法分别克隆基因片段Sj22.6ku和LHD-Sj23,以重叠延伸PCR方法将两个片段连接,构建原核重组表达质粒pET28-Sj22.6-LHD-Sj23;该重组质粒在大肠杆菌Rosetta DE3中表达后所获得的融合蛋白Sj22.6-LHD-Sj23分子量约为35 ku。Western blot鉴定结果表明该重组蛋白有良好的免疫原性,为进一步研制日本血吸虫病分子诊断试剂盒奠定了基础。
In this study,cDNA fragments of Sj22.6 ku and LHD-Sj23 gene of Schistosoma japonicum were amplified by specific primers,respectively,from mRNA of S.japonicum by RT-PCR,and fused together by overlap-extension PCR.The fusion gene was cloned into prokaryotic expression plasmid to construct pET28-Sj22.6-LHD-Sj23 and transformed to E.coli Rosetta DE3 cells.The recombinant protein approx 35 ku was expressed and its antigenicity was confirmed by western blot.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2010年第11期904-907,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
浙江省攻关项目资助
