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Analysis of Hantavirus genetic variations in suckling mouse with PCR SSCP method

Xia Guoqing 1, Liu Yanfang 1, Yang Shoujing 1, Yan Peisong 1, Feng Lei 2 1Department of Pathology, 2Central Laboratory, Fourth Military Medical University, Xi'an 710033  
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.
【Fund】: 国家自然科学基金
【CateGory Index】: R365.51
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