目的 :表达恙虫病东方体 (Orientiatsutsugamushi)Karp株 5 6 0 0 0表面蛋白 ,探索其作为诊断抗原的可能性。方法 :采用PCR方法 ,从Ot.Karp株菌种基因组DNA中扩增出 5 6 0 0 0成熟蛋白基因片段 ,将该片段克隆于原核表达载体pQE30 ,构建重组质粒pQE30 / 5 6 ,转化大肠杆菌 ,经IPTG诱导表达、SDS_PAGE电泳和免疫印迹检测目的蛋白的表达。结果 :(1)获得长约 15 0 0bp的Ot.Karp株 5 6 0 0 0外膜蛋白基因 ;(2 )SDS_PAGE电泳和免疫印迹显示该重组质粒转化的大肠杆菌有一相对分子质量约为 5 6 0 0 0的独特蛋白带 ;(3)表达后菌体超声破碎后 ,目的蛋白主要以包涵体形式存在 ;(4)重组表达质粒序列分析结果显示 ,pQE30 / 5 6中插入的基因片段的序列与已报道的Ot.Karp株5 6 0 0 0外膜蛋白基因序列基本一致。结论 :获得了Ot.Karp 5 6 0 0 0蛋白基因 ,并在大肠杆菌中实现了表达 ,表达的重组蛋白具有免疫反应性。 OBJECTIVE: To express 5600 surface protein of Karp strain Orientiatsutsugamushi and to explore its potential as a diagnostic antigen. Methods: The mature protein gene fragment of 5600 was amplified by PCR from the genomic DNA of Ot.Karp strain. The fragment was cloned into the prokaryotic expression vector pQE30 and the recombinant plasmid pQE30 / 5 6 was constructed and transformed into Escherichia coli , Induced by IPTG expression, SDS_PAGE electrophoresis and Western blot to detect the expression of the target protein. (2) SDS-PAGE electrophoresis and Western blotting showed that the recombinant plasmid transformed E. coli has a relative molecular mass of about 5 6 (4) The recombinant plasmid sequence analysis showed that the sequence of the inserted gene fragment in pQE30 / 5 6 It is basically consistent with the reported gene sequence of outer membrane protein of Ot.Karp strain 5600. Conclusion: The Ot.Karp 5600 protein was obtained and expressed in E. coli. The expressed recombinant protein was immunoreactive.