摘要
目的 :开发简单、可靠的 HTN、SEO型汉坦病毒血清学分型检测方法。方法 :应用 RT- PCR法对全 NP蛋白基因及其型特异区基因进行扩增 ,经 TA克隆及 Cui- SS、Cui- 15 5杆状病毒载体的构件、转获重组杆状病毒 ,再感染 Sf9细胞进行表达。结果 :克隆与表达了 SEO病毒 NP蛋白的全蛋白编码区及型特异编码区 (15 5到 4 2 9氨基酸处的编码基因 ) ,全 NP蛋白表达产物与病毒株感染 Vero E6细胞的反应谱完全一致 ,型特异区表达产物与相应的型特异 Mc Ab结合。结论 :建立了 SEO型病毒全 NP蛋白及 NP蛋白型特异区编码基因的克隆和表达方法 ,为开发简便、可靠的
Objective:To establish a simple and reliable method detecting SEO serotype in Hantaan virus.Methods:Total NP gene and its type specific region gene were amplified by reverse transcription polymerase chain reaction(RT-PCR).After TA clone,vector construction of Cui-SS,Cui-155 baculovirus and transposition,recombinant baculovirus were obtained.By reinfection of Sf9 cell,expression was achieved.Results:Expression product which cloning and expressing total NP coding region and type specific coding region(amino acid from 155 to 429) was in full agreement with Vero E6 cell infected by virus strain in reaction band.Expression product could combine with its type specific McAb.Conclusion:The method of clone and expression of the SEO virus antigen from NP and NP type specific region coding gene was established,and made simple and reliable measuring method of SEO serotype in Hantaan virus possible.
出处
《现代预防医学》
CAS
2002年第4期500-503,共4页
Modern Preventive Medicine
