摘要
目的：研究汉坦病毒在感染乳鼠体内有无基因变异．方法：用陈株病毒感染Ｖｅｒｏ－Ｅ６细胞并继之感染３ｄ龄乳鼠，提取病毒ＲＮＡ并反转录成ｃＤＮＡ，以ＰＣＲ分别从感染细胞和乳鼠中克隆出病毒的部分Ｇ２编码区序列，继而用非变性聚丙烯酰胺凝胶电泳对之进行单链构象多态性（ＳＳＣＰ）分析．结果：从感染细胞和乳鼠中克隆出的此位置和大小完全相同的ｃＤＮＡ片段单链电泳构像明显不同，从感染乳鼠中克隆的基因片段单链电泳速率明显快于从感染细胞中克隆的相应基因片段．结论：汉坦病毒的基因序列可因寄生宿主的不同而发生变异．
Objective: To investigate whether Hantavirus underwent genetic alterations in infected suckling mice. Methods: Vero E6 cell line monolayers were infected with Hantavirus Chen strain for the virus propagation and 3 day old suckling mice were subsequently infected with the virus in culture medium supernatant. Virus RNAs were respectively extracted from infected cells and mice by acid guanidium thiocyanate phenol chloroform method. The RNAs were then reverse transcribed into cDNAs and the same parts of G2 encoding sequences of the virus were cloned individually from infected cells and mice by PCR. Single stranded DNA conformation polymorphisms of the cloned genes were analysed by the method of non denaturing polyacrylamide gel electrophoresis. Results: The single stranded DNA electrophoretic maps of the genes cloned from infected suckling mice and cells were apparently different. Electrophoretic rate of the single strand DNA of the gene cloned from infected mice was obviously quicker than the correspondent segment cloned from infected cells. Conclusion: Hantavirus could undergo direct genetic changes owing to different hosts.
出处
《第四军医大学学报》
1997年第3期247-249,共3页
Journal of the Fourth Military Medical University
基金
国家自然科学基金
关键词
汉坦病毒
肾综合征出血热
聚合酶链反应
Hantavirus hemorrhagic fever with renal syndrome single stranded conformation polymorphism polymerase chain reaction
