流产嗜性衣原体1B株pomp18基因β折叠区的克隆与表达

为探索POMP18蛋白的生物学功能、揭示其在流产衣原体病诊断和防治中的作用,本试验依据流产嗜性衣原体S26/3株全基因序列设计引物,用PCR法扩增流产衣原体pomp18基因β折叠区,将特异性扩增片段克隆入pMD18-T载体,测序比较分析序列后确认为目的基因,提交GenBank(Accession EU326104).与GenBank中流产嗜性农原体S26/3株的pomp18基因和氨基酸序列的相似性均达99%,与其他各型衣原体代表株的相似性为79%～80%.将pomp18基因β折叠区亚克隆到pET-32a(+)表达载体上,重组表达载体转化BL21(DE3)表达宿主菌,以IPTG诱导该重组菌,结果显示该重组菌表达了59 kD大小的融合蛋白;用羊流产嗜性衣原体多克隆抗体对表达的重组蛋白进行Western blotting试验,结果显示该重组蛋白具有免疫原性.本研究首次克隆、表达了流产衣原体pomp18基因β折叠区,原核表达获得了59 kD重组蛋白,为进一步探讨其生物学功能奠定基础.

Cloning and prokaryotic expression of beta-domain of pomp18 gene of Chlamydophila abortus 1B strain

The beta-domain gene of pomp18 gene was amplified by PCR method based on the whole gene sequence of the C.abortus S26/3 strain,then the target gene was cloned into pMD18-T vector,identified by sequencing analysis and submitted to GenBank(Accession EU326104).The cloned target gene and deduced peptide sequence were compared to the beta-domain gene sequences of the characterized C.abortus S26/3 strain,they were found to be 99% and 99.0% identical to that of C.abortus S26/3 strain.Compared to other C.abortus st...