Construction of Prokaryotic Expression Vectors Harboring HV S and S Partial Gene Segments and Identification of Expressed Nucleocapsid Protein

LI Guang—yu; HUANG Chang—xing; PAN Lei; MOU Dan —lei; LI Xing—hong; ZAN G Yin; YANG Wei—song; BAI Xue—fan

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October 2003

LI Guang—yu, HUANG Chang—xing, PAN Lei, MOU Dan —lei, LI Xing—hong, ZAN G Yin, YANG Wei—song and BAI Xue—fan. Construction of Prokaryotic Expression Vectors Harboring HV S and S Partial Gene Segments and Identification of Expressed Nucleocapsid Protein[J]. Virologica Sinica, 2003, 18(5): 428-431.

Citation: LI Guang—yu, HUANG Chang—xing, PAN Lei, MOU Dan —lei, LI Xing—hong, ZAN G Yin, YANG Wei—song, BAI Xue—fan. Construction of Prokaryotic Expression Vectors Harboring HV S and S Partial Gene Segments and Identification of Expressed Nucleocapsid Protein .VIROLOGICA SINICA, 2003, 18(5) : 428-431.

汉滩病毒核蛋白羧基端多肽原核表达载体的构建及表达

1.
第四军医大学唐都医院感染病诊疗中心，陕西西安，710038

摘要

本文为建立分型检测方法，探讨了汉滩病毒(Hantaan virus，HTNV)核蛋白羧基端多肽的抗原性。首先，分别构建编码HTNV核蛋白及其羧基端多肽的原核表达载体pRSETA—S、pRSETA—S—C；然后，将其转化入表达菌 BL21(DE3)pLySs诱导表达，采用SDS—PAGE、Western．blot进行鉴定。我们成功构建了pRSET A—S及pRSET A—S—C 原核表达载体。SDS-PAGE显示目的蛋白大量表达，呈不溶状态，Western．blot显现目的蛋白具有良好的抗原性。为大量制备分型用核蛋白多肽抗原创造了条件
- 汉滩病毒(Hantaan virus
- , HTNV)：原核表达：多肽抗原
- , 核蛋白

Construction of Prokaryotic Expression Vectors Harboring HV S and S Partial Gene Segments and Identification of Expressed Nucleocapsid Protein

1.
Department ofInfectious Diseases，Tangdu Hospital,Fourth Military Medical University,X an 710038，China

Abstract

Abstract：To study the antigenicity of prokaryotic expressed products of Hantavirus(HV)S partial gene(502—1326bp)an d obtain data for serotyping antigen，the prokaryotic expression vectors pRSET A—S，pRSET A—S—C were constructed respectively．The recombinant expression plasmids were fans— formed into BL2 I(DE3)pLySs．Th e expression product were analyzed by SDS—PAGE and Western—blot． Th e results showed that the proteins were expressed at high level respectively an d could be recognized by patients’(infected by HTNV)sera distinctly and peculiarly．Th e truncated NP(aa155 to 429)could be considered as serotyping an tigen．
- Hantaan virus(HTNV)
- , Prokaryotic expression
- , Multipeptide an tigen

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Construction of Prokaryotic Expression Vectors Harboring HV S and S Partial Gene Segments and Identification of Expressed Nucleocapsid Protein

1. Department ofInfectious Diseases，Tangdu Hospital,Fourth Military Medical University,X an 710038，China

Abstract: Abstract：To study the antigenicity of prokaryotic expressed products of Hantavirus(HV)S partial gene(502—1326bp)an d obtain data for serotyping antigen，the prokaryotic expression vectors pRSET A—S，pRSET A—S—C were constructed respectively．The recombinant expression plasmids were fans— formed into BL2 I(DE3)pLySs．Th e expression product were analyzed by SDS—PAGE and Western—blot． Th e results showed that the proteins were expressed at high level respectively an d could be recognized by patients’(infected by HTNV)sera distinctly and peculiarly．Th e truncated NP(aa155 to 429)could be considered as serotyping an tigen．