PAN Lei, BAI Xue—fan, HUANG Chang—xing, LI Guang-yu．CHEN Hong—mei, W EI San—hua．YANG W ei—song, Construction of HV S and 5’Terminal of S Gene Segment Eukarytic Expression Vectors and Transfection into Vero Cells[J]. Virologica Sinica, 2002, 17(1): 47-50.
Citation:
PAN Lei, BAI Xue—fan, HUANG Chang—xing, LI Guang-yu．CHEN Hong—mei, W EI San—hua．YANG W ei—song, .
Construction of HV S and 5’Terminal of S Gene Segment Eukarytic Expression Vectors and Transfection into Vero Cells .VIROLOGICA SINICA, 2002, 17(1)
: 47-50.
汉滩病毒s基因及其5’端真核表达载体的构建及 在细胞中的表达
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第四军医大学唐都医院传染科 西安710038
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摘要
摘要：体外研究设濉病毒(HTNV)S基因及其5’端表达的意义，为核蛋自T细胞表位的研究奠定基础。设计2套 引物．用PCR方法从PBV220一$22原核质粒中扩增出s基因全读码框(37—1326bp)及s基因5’端(37—501bp)．用TA 克隆将其克隆A pcDNA3 l／V5一His~TOPO载体中．成功构建peDNA3 1-S及pcDNA3 1-S-N真核表达载体，并通过 脂质体转染至Vem细胞中，进行了瞬时表达。间接免疫荧光成功检测到pcDNA3 1-S及pcDNA3．1-S-N在Vero细 胞中的表达。pcDNA3 1-S及pcDNA3．1-S-N真核表达载体有较高的转染效率．目的基因能在宿主细胞中表达．有 利于研究HTNV—S基因在T细胞表位研究中的意义
Construction of HV S and 5’Terminal of S Gene Segment Eukarytic Expression Vectors and Transfection into Vero Cells
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Department of infectious Disease, Tangdu Hospital,The 4th．Military Medical University．Xi'an 710038,China
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Abstract
Abstract：In order tO provide some data for study of T cell epitope．HV S and 5’termirmI of S gene segment were inserted into an eukarytic expression vector pcDNA 3．1／V5一His TOPO．The recombi— nant expression plasmids peDNA3 1-S and pcDNA3．1-S—N were constructed．Vero cells were trans— fected in vitro with them．The transient expresion of HV S and 5’terminal of S gene segment in veto cells w∞ detected by；ndirect immunofluoreseence assay(IFA)．
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Proportional views
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