CAJ | 학술논문

采用RT-PCR方法从河南地区甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)感染的玉米叶片中,克隆了辅助成分-蛋白酶基因(helper component.proteinase,HC-Pro).该基因长1 380 bp,编码460个氨基酸.序列比对表明.HC-Pro编码的氨基酸序列与北京、山东地区SCMV分离物的相似性分别达到97%和98%.而与以前数据库登录的河南分离物的相似性仅98%,说明河南地区SCMV可能发生了变异.选取长582 bp的HC-Pro片段,构建了表达发夹RNA的基因沉默双元表达载体pRNAi-HC-Pro(±),成功转化了农杆菌LBA4404.
채용RT-PCR방법종하남지구감자화협병독(Sugarcane mosaic virus,SCMV)감염적옥미협편중,극륭료보조성분-단백매기인(helper component.proteinase,HC-Pro).해기인장1 380 bp,편마460개안기산.서렬비대표명.HC-Pro편마적안기산서렬여북경、산동지구SCMV분리물적상사성분별체도97%화98%.이여이전수거고등록적하남분리물적상사성부98%,설명하남지구SCMV가능발생료변이.선취장582 bp적HC-Pro편단,구건료표체발협RNA적기인침묵쌍원표체재체pRNAi-HC-Pro(±),성공전화료농간균LBA4404.
The complete helper component-proteinase (HC-Pro) gene was cloned by RT-PCR from diseased maize leaves infected by Henan isolate of Sugarcane mosaic virus (SCMV).The eDNA of the HC-Pro contains a 1 380 bp open reading frame (ORF) and encodes 460 amino acids.Sequence alignment showed that the deduced amino acid sequence of the HC-Pro shared 97% and 98% similarities to homologues from SCMV Beijing isolate and Shandong isolate respectively.However, the He-Pro cloned by our group only shared 98% similarity to that of SCMV Henan isolate deposited in the Genbank, and this might indicate that the SCMV Henan isolate has undergone some genetic divesities.A 582 bp long fragment of the HC-Pro was selected to construct the binary expression vector pRNAi-HCPro (+) with hairpin RNA structure, and the vector was successfully transformed into the agrobacterium strain LBA4404.