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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 ,并就干扰素α2b抑制SARS病毒的可能分子机制及其潜在的应用价值进行了讨论
Objective To investigate the molecular mechanism of recombinant interferon α2b inhibiting the replication of SARS virus at the cellular level by using the genome-wide chip technology of SARS virus. Methods According to the published sequence of SARS virus genome and bioinformatics software, PCR primers were designed to include all possible coding regions of SARS virus genome. All the PCR products were used to prepare SARS virus genome-wide cDNA microarray. The chip was used to investigate the molecular basis of recombinant interferon α2b inhibiting SARS virus replication. Results The whole genome of SARS virus cDNA was developed by PCR and the cDNA microarray was used to detect the RNA level of each SARS virus. Using this technique, the molecular mechanism of interferon against SARS virus has been studied. The results showed that recombinant human interferon α2b significantly inhibited the transcription of almost all viral RNAs during SARS virus replication and showed a dose-dependent relationship. A new gene of SARS virus with no definite function was found, which was named as U gene and transcribed The earliest, highest transcription level, is also very sensitive to the inhibitory effect of interferon; it may play an important role in the transcriptional replication of the virus. Conclusion The SARS coronavirus cDNA microarray can be used to study the molecular mechanism of virus replication and related anti-viral drugs including interferon. In this paper, the transcriptional activity of human interferon alpha 2b inhibiting the replication of SARS virus was studied at the molecular level, and the possible molecular mechanism of interferon alpha 2b inhibiting SARS virus and its potential application were discussed