南方水稻黑条矮缩病毒SP7-1植物表达载体的构建及遗传转化  

Construction and genetic transformation of plant expression vector of southern rice black-streaked dwarf virus SP7-1

作　　者：苑侠[1,2] 孙枫[2] 卢嫣红[2] 周彤[2] 范永坚[1,2] 周益军[2] 

机构地区：[1]南京农业大学,农业部病虫害监测与治理重点开放实验室,江苏南京210095 [2]江苏省农业科学院植物保护研究所,江苏省植物病毒病诊断检测中心,江苏南京210014

出　　处：《江苏农业学报》2012年第6期1278-1282,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家公益性行业(农业)科研专项(201003031);江苏省自然科学基金(BK2010018);国家"973"计划(2010CB126203);江苏省农业科技自主创新基金项目[CX(10)207]

摘　　要：克隆南方水稻黑条矮缩病毒(Southern rice black-streaked dwarf virus,SRBSDV)的SP7-1基因,构建植物表达载体并转化拟南芥,为研究SP7-1致病机理提供基础。根据已发表的SRBSDV SP7-1序列,设计特异性引物,通过RT-PCR的方法从感病水稻植株中克隆到完整的SP7-1基因。通过酶切连接,构建了SP7-1植物表达载体pCHF3/SP7-1,通过农杆菌介导的花序浸润法转化野生型拟南芥,获得转化植株T0代种子。对收获的T0代种子进行卡那霉素筛选,获得了4株阳性植株。阳性植株的PCR和RT-PCR分析检测结果表明,构建的SP7-1载体已成功转化拟南芥,病毒SP7-1已经整合至拟南芥基因组中并能正常表达。The full-length SRBSDV（southern rice black-streaked dwarf virus）SP7-1 gene was cloned from diseased rice by RT-PCR using a primer designed according to the NCBI GenBank（accession number：EU784841）.SP7-1 plant expression vector pCHF3/SP7-1 was constructed,and were transformed into wild type Arabidopsis thaliana（Col-0）by floral dip method.All four positive transgenic seedlings were identified from T0 transgenic seeds through kanamycin screening.The results of PCR and RT-PCR showed that SRBSDV SP7-1 gene was integrated into Arabidopsis genome and transcriptional level was higher than that of no-transgenic seedlings.The SRBSDV SP7-1 transgenic Arabidopsis seedlings might provide material for further studies on the functions of SP7-1.

关 键 词：南方水稻黑条矮缩病毒 SP7-1基因 拟南芥 转化 

分 类 号：Q78[生物学—分子生物学]