月季表观修饰因子DRM3基因沉默载体的构建与作用方式  

The Construction and Action of the Gene Silencing Vector of the Epigenetic Modifier Genes DRM3

作　　者：韩凯 董然[1] 李永红[2] HAN Kai;DONG Ran;LI Yonghong(Jilin Agricultural University,Changchun,Jilin 130118;Shenzhen Polytechnic,Shenzhen,Guangdong 518055,China)

机构地区：[1]吉林农业大学,吉林长春130118 [2]深圳职业技术学院,广东深圳518055

出　　处：《深圳职业技术学院学报》2018年第5期47-51,共5页Journal of Shenzhen Polytechnic

基　　金：国家自然科学基金资助项目(601523K27028)

摘　　要：利用Rose Transcriptome Database获取月季DRM3片段,根据所获取的目的序列分别设计一对添加了酶切位点的引物DRM3-Xba I-F和DRM3-Kpn I-R,利用反转录得到的c DNA为模板进行扩增,得到DRM3基因沉默片段,再将酶切的片段连接至T4载体,连接产物转化大肠杆菌DH5α.酶切和序列分析鉴定表明扩增得到的重组质粒正确,命名为pTRV2-DRM3.利用构建好的载体将月季扦插小苗进行病毒诱导的基因沉默(VIGS),获得DRM3沉默植株.其结果为研究低温影响月季花器官发育的网络调节提供基础.Using Rose Transcriptome Database to get Chinese Rose DRM3 fragments, a pair of primers added enzyme loci DRM3 XbaI - F and DRM3 KpnI - R is designed respectively based on the sequence of purpose. A reverse transcription of eDNA as a template for amplification is used to acquire DRM3 fragment gene silencing. Then enzyme fragment is connected to the T4 carrier and connecting product is turned into E.coli DH5α. Enzyme digestion and sequence analysis shows that the amplified recombinant plasmid is correct and hence named pTRV2-DRM3. The virus-induced gene silencing （VIGS） is carded out by using the constructed vector to cut the seedlings in the Chinese rose and get the silencing plant of DRM3 gene. The result provides a theoretical basis for studying the regulating network of flower organ development under low temperature.

关 键 词：月季 载体构建 病毒诱导的基因沉默 DRM3 

分 类 号：Q943.2[生物学—植物学]