MA Zhong—liang, YANG Huai—yi and TIAN Bo*. Expression of Non—structural Protein S6 of Rice dwarf virus in E coil and Preparation of Its Antiserum[J]. Virologica Sinica, 2003, 18(5): 496-499.
Citation:
MA Zhong—liang, YANG Huai—yi, TIAN Bo*.
Expression of Non—structural Protein S6 of Rice dwarf virus in E coil and Preparation of Its Antiserum .VIROLOGICA SINICA, 2003, 18(5)
: 496-499.
水稻矮缩病毒非结构蛋白基因 6的表达和抗血清的制备
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摘要
摘要:绎RT-PCR扩增⋯水稻矮缩病毒(RDV)中国分离物非结构蛋白基因s6,并克隆至pGEM-Teasy载体上。 序列分析表明该基因与日本株具有高度同源性,并且含有较高比例的稀有密码子。将s6基因克隆到表达载体 pGEX一6P—l,并转化大肠杆菌,该基因在大肠杆菌以包涵体形式大量表达。以表达的融合蛋白作抗原免疫家兔, 制备抗S6 白的抗血清,ELISA 测定表明,该血清与抗原共价特异件反应,抗血清的效价为1:3000.Westernblot 印迹实验表明该抗血清能特异性榆测RDV感染的水稻组织中的s6蛋白,因而可作为感染RDV的水稻植株的分 子手段。
Expression of Non—structural Protein S6 of Rice dwarf virus in E coil and Preparation of Its Antiserum
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1.
Department of Molecular Virologogy,Institue ofMicrobiology,Chinese Academy of Sciences,Beijing 100080,China
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Abstract
Abstract: gene of a Chinese isolate of Rice dwarf virus fRDV)was amplified by RT.PCR and then cloned into pGEM—Teasy.Sequence analysis showed that its nucleotide sequence had high identity to a Japan ese isolate of RDV an d contmned high percent of rare codons. l6 gene was expressed m inclusion body with high yield in E.coli after being subcloned into expression vector pGEX一6P—1.Immune rabbit witll S6 protein obtained the antiserum.The an tiserum having a titer of l:3000 tested by ELISA Can specifically react wim the expressed S6 protein.Our study confn-med that the an tiserum could be used to detect S6 protein in RDV-infected rice tissues.
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References
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Proportional views
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