5′UTR序列对DR5::GUS基因瞬时表达的影响  

Different 5′UTR sequences influence the transient expression of DR5::GUS

作　　者：王亚红[1] 赵燕[1] 彭彦[1] 刘晓柱[1] 张学文[1] 

机构地区：[1]湖南农业大学生物科学技术学院,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2012年第2期146-149,共4页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：湖南省教育厅项目(SCX1103)

摘　　要：利用生长素响应原件(DR5)构建了DR5::GUS报告基因表达载体,并在载体构建时,对GUS的5′UTR序列进行2种设计:一种是利用载体GUS基因原有的5′UTR构建成pBI121-DR5::GUS;另一种采用植物增强表达的常用原件烟草花叶病毒(TMV)的5′端序列(Ω′)替换GUS基因原有的5′UTR序列,构建成pBI121-DR5(Ω′)::GUS。将2种载体转化根癌农杆菌后,采用烟草叶片注射浸染的瞬时表达检测法对2个载体的植物表达效果进行检测,2种载体分别转化烟草叶片2 d后,对浸染的叶盘进行GUS染色分析,pBI121-DR5::GUS的转化叶片有明显的GUS反应,而pBI121-DR5(Ω′)::GUS的转化叶片则无GUS反应。分离注射浸染部位叶盘RNA后,对GUS特异引物进行RT-PCR分析,2种转化的基因都已有效转录出RNA,但前者能翻译出相应的Ω-葡萄糖苷酸酶,后者则不能完成翻译过程,说明Ω′序列的引入并未有效提高GUS基因的表达,反而抑制了mRNA的翻译。Using auxin-response element（DR5）,we constructed reporter expression plasmids containing the recombinant reporter gene DR5：：GUS for study of the auxin distribution in vivo.Two constructions were designed using pBI121,one of which is pBI121-DR5（Ω′）：：GUS,where the 5′UTR of GUS was replaced by the 5′UTR of TMV（Ω′） and the other is pBI121-DR5：：GUS with 5′UTR of GUS unchanged.We transferred the two recombinants into tobacco leaf by infiltration and the transient expression was tested two days later.The GUS reaction was strong with infiltrated pBI121-DR5：：GUS but there was no GUS activity with the pBI121-DR5（Ω′）：：GUS.RNA was isolated from the infiltrated leaf discs and subjected to RT-PCR with specific primers of GUS.The GUS mRNAs were detectable in the infiltrated cells transferred with either pBI121-DR5：：GUS or pBI121-DR5（Ω′）：：GUS,and the corresponding β-glucosidase expression was achieved in the former plasmid but not the latter plasmid.It recommended the 5′UTR sequence of Ω′ do not improve the gene expression of GUS,but do inhibit the translation of the mRNA in our study.

关 键 词：5′非编码区 生长素基因报告系统 瞬时表达 

分 类 号：Q786[生物学—分子生物学]