肺炎嗜衣原体MOMP基因的克隆与原核表达

Cloning and Prokaryotic Expression of the MOMP Gene of Chlamydophila Pneumoniae

克隆了肺炎嗜衣原体外膜主蛋白(major out membrane protein,MOMP)基因,并进行了原核表达。根据GenBank公布的肺炎嗜衣原体MOMP基因序列,设计克隆引物后用普通PCR方法扩增出肺炎嗜衣原体MOMP基因的完整片段,将其克隆到pMD-19T后进行测序,经序列比对正确后将此片段亚克隆到表达载体pET-32a(+)中,转化至大肠杆菌Rosetta(DE3),经IPTG诱导表达,再采用SDS-PAGE和Western-Blot检测重组蛋白的表达情况。结果表明:本研究克隆了肺炎嗜衣原体MOMP基因并在原核系统中成功表达了MOMP重组蛋白,且WesternBlot显示该重组蛋白具有免疫学活性。

The MOMP complete gene of chlamydophila pneumoniae was cloned and expressed in pro- karyotic expression system. The MOMP complete gene of Chlamydophila pneumoniae was amplified by normal PCR with designed cloning primers based on the reported in GenBank. The amplified MOMP complete gene was inserted into the cloning vector pMD19-T and then was sequenced. The correct MOMP gene was sub-cloned into expressed vector pET-32a（ ＋ ）for prokaryotic expression in Rosetta （DE3） induced by IPTG. The MOMP gene was cloned and was submitted to NCBI. The recombinant proteins were expressed successfully in prokaryotic expression system. Western-Blot showed that the recombinant protein has antigenicity.