大麦黄矮病毒PAV株系的CP与GFP融合基因的构建和表达  

Constructing and expressing of the CP of BYDV-PAV and GFP fusion gene in Escherichia coli

作　　者：孙润红[1,2,3] 吴云锋[2] 张银武 魏婷[2] 武科科[2] 赵震[1] 杨洋[1] 

机构地区：[1]西北农林科技大学生命科学学院,杨凌712100 [2]西北农林科技大学植物保护学院,陕西省农业分子生物学重点实验室,杨凌712100 [3]河南驻马店上蔡县农技站,上蔡463800

出　　处：《植物病理学报》2010年第5期548-551,共4页Acta Phytopathologica Sinica

基　　金：公益性行业科研专项(nyhyzx07-051);转基因抗病毒小麦新品种培育(2009ZX08002-003B);高等学校学科创新引智计划资助项目(B07049)

摘　　要：According to the published nucleotide sequences,CP gene of barley yellow dwarf virus PAV(BYDV-PAV) was synthesized by reverse transcription polymerase chain reaction(RT-PCR).The reporter gene GFP was put to the downstream of whole CP gene sequence of BYDV-PAV after double enzyme digestion,PCR identification and sequencing.The green fluorescence protein expression system of coat protein of BYDV-PAV was successfully constructed and expressed in Escherichia coli BL21(DE3)plysS.The expression product was confirmed by SDS-PAGE,Western blot analysis and fluorescence microscope.The fusion gene CP-GFP was expressed in the range of 15-22℃,but could not be expressed at 23-37℃.The results provided a base for studying on BYDV-PAV movement in aphid further.According to the published nucleotide sequences,CP gene of barley yellow dwarf virus PAV（BYDV-PAV） was synthesized by reverse transcription polymerase chain reaction（RT-PCR）.The reporter gene GFP was put to the downstream of whole CP gene sequence of BYDV-PAV after double enzyme digestion,PCR identification and sequencing.The green fluorescence protein expression system of coat protein of BYDV-PAV was successfully constructed and expressed in Escherichia coli BL21（DE3）plysS.The expression product was confirmed by SDS-PAGE,Western blot analysis and fluorescence microscope.The fusion gene CP-GFP was expressed in the range of 15-22℃,but could not be expressed at 23-37℃.The results provided a base for studying on BYDV-PAV movement in aphid further.

关 键 词：大麦黄矮病毒 PAV 融合基因 株系 GFP 病毒性病害 黄症病毒科 CP 

分 类 号：S435.121.4[农业科学—农业昆虫与害虫防治]