摘要
【目的】克隆猪流产嗜性衣原体青海株主要外膜蛋白(Major outer membrane protein,MOMP)基因,并进行序列分析及原核表达。【方法】根据GenBank公布的猪流产嗜性衣原体MOMP基因的核苷酸序列,设计并合成4条特异性引物,用套式PCR方法扩增猪流产嗜性衣原体青海株MOMP基因,将其克隆入pMD18-T载体中,进行测序及序列分析。然后将MOMP基因亚克隆入原核表达载体pGEX4T-1中,在大肠杆菌BL21(DE3)中用IPTG诱导表达,对表达产物进行SDS-PAGE和Western blot检测。【结果】扩增到猪流产嗜性衣原体青海株1170bp的MOMP全长基因。序列分析结果表明,该基因与已发表的猪流产嗜性衣原体B11001株和CP/12株核苷酸同源性均为99.7%。SDS-PAGE电泳可检测到分子质量约为66ku的融合蛋白,主要以包涵体形式存在。Western-blot分析表明,重组蛋白可被猪流产嗜性衣原体抗体识别。【结论】成功克隆了猪流产嗜性衣原体青海株MOMP基因,并进行了原核表达,表达的蛋白具有抗原活性。
【Objective】The outer membrane protein(MOMP)gene of Swine Chlamydophila abortus(CA)Qinghai strain was cloned,analyzed and expressed in E.coli.【Method】According to the published complete nucleotide sequence of CA in GenBank,four primers specific to the full-length of MOMPgene were designed.The MOMPgene fragment amplified by nested PCR was cloned into pMD18-Tvector.The recombinant plasmid was sequenced and MOMPgene was compared with other GeneBank s CA strains.MOMPgene segment was amplified from recombinant pl...
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2010年第7期33-38,43,共7页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家科技基础性工作专项(2008FY210200)
陕西省科技攻关项目(2009K02-01)
西北农林科技大学"青年学术骨干"支持计划项目(E111020901)
