梅毒螺旋体重复蛋白K的克隆表达

Cloning and expression of Treponema pallidum repeat protein K

目的体外克隆并表达梅毒螺旋体重复蛋白K,为制备预防性疫苗奠定基础。方法利用PCR反应及基因重组技术,扩增重复蛋白K编码基因,构建重组质粒pET28b㈩/TprK,IPTG诱导表达。结果PCR法扩增出了1350bp的目的片段,原核表达质粒pET28b㈩/TprK酶切鉴定正确,测序结果与Gengbank上公布的该基因序列完全一致,并可在大肠杆菌中高效表达。结论成功构建人pET28b㈩/TprK原核表达载体,并能高效表达TprK蛋白,为梅毒的预防性疫苗的制备以及进一步研究其血清学诊断试剂奠定了基础。

Objective To clone and in vitro express TprK and provide a good foundation for preparing prophylactic vaccine. Methods PCR and gene recombinant methods were used and the gene encoding TprK was amplified and inserted into expression vector pET28b（＋） after T-A cloning,. The recombinant plasmid was induced by IPTG to express TprK protein. Results The target gene at a length of 1350bp was amplified. Restriction map proved that recombinant plasmid pET28b（＋）/ TprK was correctly constructed. The sequence of target gene inserted into the recombinant plasmid was completely identical to that reported in GenBank, and efficiently expressed TprK protein in E. coli DH 5 . Conclusion Prokaryotic expression vector pET28b（＋）/TprK has been constructed successfully, and pET28b（＋）/TprK protein expressed to have set foundation for preparing prophylactic vaccine and developing serologically diagnostic kit for TP infection.