10株鹦鹉热衣原体菌株主要外膜蛋白基因的比较性研究

Comparative study on the major outer-membraneprotein gene of ten isolates of Chlamydia psittaci

目的 进行比较性研究了 10株鹦鹉热衣原体菌株 ,分析其主要外膜蛋白 (MOMP)基因的限制性内切酶图谱和基因同源性。方法 接种鸡胚卵黄囊对菌株进行培养 ,收集培养物 ,通过聚合酶链式反应 ,扩增了 10株Cps的主要外膜蛋白基因 ,获得 10 5 8bp的片段 ,根据限制性片段长度多态性 (RFLPs)技术 ,利用AluⅠ酶切此片段 ,获得其限制性图谱。将此片段连入 pGEMT载体 ,进行基因测序。 结果 有 5株菌的MOMP基因完全相同 ,将它们与其它 5株比较 ,仅有个别碱基不同。

Aim To reveal the homology of the major outer-membrane protein(MOMP) gene of ten isolates of Chlamydia psittaci,and study on the restriction endonuclease map and nucleotide sequences.Methods Ten isolates of Chlamydia psittaci,obtained from five different hosts,were cultured in embryonated chicken eggs.On embryo death,yolk sacs were harvested,and preparations of DNA of the isolates were compared by analyzing Alu I restriction of MOMP gene after DNA amplification by the polymerase chain reaction(PCR).Then the productions of PCR were linked into pGEM T vector and sequenced.Results A strict correspondence is observed between the MOMP gene restriction pattern of the ten isolates.After sequenced,five of the isolates are identical,and only a few basepairs were different to the other five straias.The results indicate that the MOMP gene is a highly stringent gene in Chlamydia psittaci.