水稻条纹病毒编码的NS2蛋白的亚细胞定位分析

Subcellular localization analysis of NS2 encoded by Rice stripe virus

通过RT-PCR克隆获得水稻条纹病毒(RSV)编码的蛋白NS2和拟南芥柯浩体蛋白Atcoilin的cDNA,将两者分别与表达载体pEarley102(带青色荧光蛋白,CFP)和pEarley101(带黄色荧光蛋白,YFP)同源重组.将构建好的重组载体分别转化到农杆菌EHA105中,通过农杆菌接种法将两者共同注射到本氏烟叶片表皮细胞中进行表达.在共聚焦显微镜下观察,NS2-CFP与Atcoilin-YFP能重叠在一起,表明RSV NS2定位于柯浩体.

The cDNAs of NS2 encoded by Rice stripe virus and the Arabidopsis thaliana coilin （Atcoilin） were amplified by RT- PCR, and were inserted individually into expression vector pEarley102 bearing cyan fluorescent protein （CFP） and pEarleyl01 bearing yellow fluorescent protein （YFP）. Recombinant proteins were transiently expressed on the leaf epidermis cells of Nicotiana benthamiana by using Agro-infiltration. Under the confocal laser scanning microscope, we determined that NS2-CFP and Atcoilin- YFP were colocalized. The data suggested that the RSV NSE was co-localized with cajal bodies.