病毒诱导的基因沉默技术实验教学设计和实践  

Experimental design and practice for plant gene function analysis by VIGS

作　　者：侯巧明[1] 张立[1] 贺新强[1] 丁琦[1] 

机构地区：[1]北京大学生命科学学院生物基础教学实验中心,北京100871

出　　处：《实验技术与管理》2015年第9期191-193,共3页Experimental Technology and Management

摘　　要：病毒诱导的基因沉默(VIGS)技术是近年发展起来研究植物基因功能的新方法。利用含有目的基因片段的病毒载体侵染植物,导致植物体内相应的功能基因表达受到抑制成为沉默基因,相应功能缺失,从而推测目的基因的功能。该文克隆了烟草番茄红素脱氢酶(PDS)基因片段,插入到烟草脆裂病毒载体(TRV)中,构建了pTRV-PDS的VIGS载体,转入农杆菌侵染烟草后成功沉默了PDS基因,使烟草叶片出现白化表型,成功建立了烟草VIGS体系。将VIGS这个当前生物学研究热点引入教学实验中,改革更新实验教学内容,有益于学生扩展科研思维。VIGS is a recently developed approach for the gene function analysis in plant .Virus vectors with target gene being transformed into the plant can result in the endogenous gene expression silent ,followed with the loss of corresponding function in plant .VIGS technology uses this mechanism to silence plant genes in order to determine their function .Phytoene desaturase （PDS） gene fragment is cloned into tobacco rattle virus vector （TRV） to obtain pTRV‐PDS .The construction is transformed into A grobacterium GV3101 and then infected N .benthamiana .Silencing of the PDS gene in N .benthamiana can result in the white leaf due to photo‐bleaching ,indicating successful establishment of the VIGS system .Introduction of VIGS ,the current hot research ,into the experimental teaching is beneficial to expanding the students’ scientific field .

关 键 词：VIGS RNAI 烟草脆裂病毒 番茄红素脱氢酶 

分 类 号：G642.0[文化科学—高等教育学]