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兔多杀性巴氏杆菌热休克蛋白60基因的原核表达及抗原性分析

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摘要 为了研究多杀性巴氏杆菌(P.multocida,Pm)热休克蛋白60(Hsp60)基因表达产物的抗原性,试验根据已发表的Pm 70株序列(AE004439)设计了1对引物,采用PCR技术扩增了兔多杀性巴氏杆菌C51-17株Hsp60的基因序列,将扩增产物与表达载体pPRO-EX-htb连接,得到重组表达质粒pHT-Hsp60,再用BamHⅠ和XhoⅠ双酶切鉴定并测序确认正确后,将重组表达质粒pHT-Hsp60转化大肠杆菌BL21(DE3)并诱导表达,进行SDS-PAGE分析和Western-blot检测。结果表明:SDS-PAGE表达蛋白约为60 ku,与预期的Hsp60分子质量一致;表达产物与兔多杀性巴氏杆菌阳性血清发生特异性反应,说明表达产物具有良好的抗原性。
出处 《黑龙江畜牧兽医》 CAS 北大核心 2012年第4期108-110,共3页 Heilongjiang Animal Science And veterinary Medicine
基金 黑龙江省科技攻关项目(PC09S02)
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