中国比较医学杂志2012，Vol.22Issue(10)：37-42,6.DOI:10.3969.j.issn.1671.7856.2012.010.009
RT-SHIV rt基因单拷贝PCR扩增方法的建立及初步应用
Establishment and Application of a Single Genome Amplification Assay for RT-SHIV rt Gene Detection
摘要
Abstract
Objective To establish a single genome amplification assay for obtaining complete rt gene sequence from RT-SHIV chimeric virus. Method Specific primers were designed with Oligo and screened through serial dilution, then PCR was optimized to amplify complete rt gene by selecting the best annealing temperature and cycles. Serial dilutions of RT-SHIV chimeric plasmid was used as template standards to establish this system. Complete rt gene of infected monkey in vivo were amplified using this method, and the obtained sequences were analyzed with BioEdit software. Results A group of nested PCR primers was got and a single genome amplification assay was established to obtain complete rt gene. Single genome sequences were obtained using dilution of 100 copies/μL plasmid. The results show that the RT-SHIV-infected monkeys had one and six amino acid variations at d266 and d294. Conclusions The single genome amplification assay established in this study is an highly sensitive, speeifie and reprodueible method. It ean be used to analyze eomplete rt gene from various types of RT-SHIV.关键词
RT-SHIV/单拷贝PCR/rt基因Key words
RT-SHIV/Single genome amplification assay/rt gene.分类
医药卫生引用本文复制引用
鞠斌,王卫,丛喆,姚南,陈霆,杜文达,苏爱华,高锡强,魏强..RT-SHIV rt基因单拷贝PCR扩增方法的建立及初步应用[J].中国比较医学杂志,2012,22(10):37-42,6.基金项目
国家科技重大专项课题(2012ZX10001-007-008和2012ZX10004-501). （2012ZX10001-007-008和2012ZX10004-501）
