汉坦病毒全NP蛋白及其型特异区编码基因的克隆及表达

Clone and Expression of Total NP and NP Type Specific Region Coding Gene of SEO Virus

目的 开发简单、可靠的HTN、SEO型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、转座、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VeroE6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McAb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SEO型血清学分型检验方法奠定了基础。

ObjectiveTo establish a simple and reliable method for measurement of SEO serotype in Hantan virus.MethodsTotal NP gene and its type specific region gene were amplified by reverse transcription polymerase chain reaction(RT-PCR).After TA clone,vector construction fo Cui-ss,Cui-155 baculovirus and transposition,recombinant baculovirus was obtained .By reinfection of Sf9 cell,expression was achieved.ResultsExpression 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.ConclusionThe 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 measurement method of SEO serotype in Hantaan virus possible.