SARS冠状病毒全基因组cDNA芯片的研制以及人重组干扰素α2b抑制SARS病毒复制分子机制的初步研究

Development of the cDNA chip for SARS virus and a primary study on the possible molecular mechanism of interferon α2b inhibiting the SARS virus replication

目的 在细胞水平上 ,利用研制的SARS病毒全基因组芯片技术探讨人重组干扰素α2b抑制SARS病毒复制的分子机制。方法 根据已经发表的SARS病毒基因组全序列和生物信息学软件分析 ,设计包括SARS病毒全基因组的各个可能编码区的PCR引物 ;将所有PCR产物用来制备SARS病毒的全基因组cDNA芯片。利用该芯片来探讨人重组干扰素α2b抑制SARS病毒复制的分子基础。结果 通过PCR方法研制了SARS病毒全基因组的cDNA芯片 ,并说明cDNA芯片可以准确地检测SARS病毒各个基因的RNA水平。利用这一技术研究了干扰素抗SARS病毒的分子机制。结果表明 ,人重组干扰素α2b可以明显抑制SARS病毒复制过程中几乎所有病毒RNAs的转录 ,并呈现一定的剂量关系 ;并发现一个目前还没有确定功能的SARS病毒新基因 ,命名为U基因 ,转录最早 ,转录水平最高 ,对干扰素的抑制作用也很敏感 ;它可能在病毒的转录复制过程中发挥重要作用。结论 SARS冠状病毒cDNA芯片可以用来研究病毒复制的分子机制以及相关抗病毒药物包括干扰素的研究。本文在分子水平上研究了人干扰素α2b抑制SARS病毒复制的基因转录动态 。

Objective To study the molecular mechanism of interferon α2b(IFNα2b) inhibiting the SARS virus replication .SARS associated coronavirus (SARSvirus)cDNA chip was developed and applied to detect the virus RNA transcrip tionlevels in the interferon treated and untreated cell cultures ,and the mechanism of anti SARS virusa ctivity of interferon α2b in cellculture system was explored .Methods SARS virusc DNA chip was prepared by comparing the published SARS virusgenome sequence,and the cDNA chip was used to study the interferon α2 bf unction during SARS virus replication .Results SARS virusc DNA chip was successfull yprepared by using PCR method .There sults showed that the cDNA chip could beused to detect the viral RNA transcription level .Interferon α2b could in hibital most allt he SARS virusgene trans cription .Anun knowngene atthepo sition 2 8130 2 84 2 6bp ,namedasUgene ,may play an important roleduring the viralre plication .Conclusion ASARS virus whole genomec DNA chip wase stablished .It could beused to study the virus molecular biology and antiviral drugs creening .There sult salso showed tha tin terfer on α2b could in hibit almost the whole virusgene transcription byusingthecDNAchip .