摘要
目的表达奶牛源鹦鹉热衣原体ompA基因,探索其作为诊断抗原的可能性。方法采用套式PCR方法扩增出了奶牛源鹦鹉热衣原体ompA基因完整片段,此扩增产物经过双酶切后克隆到表达载体pGEX-4T-1中,转化大肠杆菌BL21(DE3),经IPTG诱导表达后采用SDS-PAGE和Western-blot检测重组目的蛋白的表达结果。结果重组蛋白经奶牛衣原体阳性血清鉴定正确。结论ompA基因在原核表达系统中得到正确表达。
The ompA gene of Chlamyia psittaci in cows was amplified by PCR with primers designed based on those reported in GenBank. The amplified ompA gene was inserted into the bacterial plasmid vector pGEX-4T-1 and then transformed into E. coli BL21 （DE3） with IPTG induction. The gene was derived from plasmid pMD18-T vector and then sequenced. It was demonstrated that this recombinant fusion protein of approximately 68kD in molecular mass was highly expressed in inclusion body and more pure proteins would be produced after purification. The fusion protein specifically reacted with positive sera of bovine Chlamydia as demonstrated by Western blotting. These results indicate that this recombinant fusion protein shows good reactivity and could be used to develop the diagnostic kit for bovine Cklarnydia and genetic engineering vaccine.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2010年第2期140-143,共4页
Chinese Journal of Zoonoses
基金
国家科技支撑计划项目(2006BAD04A05)
国家科技基础性工作专项(2008FY210200)基金联合资助
