中国莱姆病螺旋体PD91外膜蛋白C表达载体的构建及序列分析

目的　构建莱姆病螺旋体PD91菌株外膜蛋白C(OspC)的表达载体 ,克隆表达OspC以便用于莱姆病诊断研究。方法　设计引物 ,用PCR从PD91扩增出ospC基因 ,定向克隆到表达载体PGEX - 5X - 1,构建重组质粒。通过PCR、酶切分析及序列测定等方法鉴定重组质粒。结果　ospC基因被正确克隆到表达载体PGEX - 5X - 1中。序列测定结果证实与国外已报道的ospC基因序列同源性在 6 5 %～ 92 %。 结论　OspC的编码基因在不同菌株间的同源性存在较大的差异。PGEX -5X - 1-ospC重组质粒的成功构建 ,为我国莱姆病特异性诊断试剂的研究奠定了基础。

Construction of an expression vector PGEX-5X-1-ospC carrying the outer surface protein C Gene from Borrelia burgdorferi PD19 strain and sequence determination

Aim To construct an expression vector carrying the B garinii major outer surface protein C(OspC) gene which was cloned and expressed for diagnoses of Lyme disease Method The ospC gene was amplified from the genome of B garinii PD91 strain by PCR and recombined with plamid PGEX 5X 1 The recombinant plasmid PGEX 5X 1 ospC was identified with PCR,restriction endonuclease analysis and sequencing Results ospC gene was cloned correctly into vector PGEX 5X 1 The homology of nucleotide sequence of the inserted fragment in the plamid was between 65% to 92% compared with the published sequence of ospC gene Conclusion The homology present highly difference among various isolates,and the PGEX 5X 1 ospC recombinant plasmid has been constructed successfully for the research on special diagnosis of Lyme disease