马铃薯X病毒山西分离物外壳蛋白基因的克隆和核苷酸序列分析  

Cloning and Sequencing of cDNA Encoding Coat Protein for Potato Virus X

作　　者：白云凤[1] 白冬梅[1] 张维锋[1] 田芙蓉[2] 

机构地区：[1]山西省农业科学院作物遗传研究所,太原030031 [2]山西省大同市农业局,山西大同037000

出　　处：《应用与环境生物学报》2008年第5期599-603,共5页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金资助项目(No.30471102);山西省国际科技合作项目(No. 2007081002);山西省回国留学人员基金项目(No. 200689);山西省科技攻关项目(No. 20080311016-2)资助~~

摘　　要：对GenBank公布的马铃薯X病毒的基因组核苷酸序列和外壳蛋白基因的核苷酸序列进行了同源性比对分析．结果表明，马铃薯X病毒外壳蛋白基凶核苷酸序列的同源性要高于基因组全序列的同源性，并且外壳蛋白基因3端核苷酸序列的同源性又高于5＇端．设计一对特异性引物，以马铃薯感病植株的总RNA为模板，经RT-PCR扩增得到了马铃薯X病毒外壳蛋白基因．该基因含714个核苷酸，编码238个氨基酸．核苷酸序列比对结果表明，该基因与Genbank公布的马铃薯X病毒其他分离物的外壳蛋白基因的同源性为80．2％～97．5％，且3’端的同源性要高于5’端．上述结果预示，RNA干扰抗病毒基因工程中，利用马铃薯X病毒外壳蛋白基因的3’端比用5’端是更好的策略．Eight available genomic complete nucleotide sequences and coat protein gene （cp） nucleotide sequences of potato virus X （PVX） isolates reported in GenBank were analyzed and compared. It was showed that cp nucleotide sequences shared higher identity than complete nucleotide sequences among PVX isolates. A pair of specific primers was designed based on PVX genomic sequences reported in GenBank, and cDNA （714 bp long） encoading cp gene of PVX was amplified by reverse transcription polymerase chain reaction （RT-PCR） using the total RNA extracted from susceptible potato plants as a template. The nucleotide sequence of the cloned cp gene was compared with that of homologous gene of all the above eight available PVX isolates. It was revealed that the identities ranged from 80.2% to 97.5% and the 3-terminal part of cp gene shared higher identity than 5-terminal part in nucleotide sequence. It was indicated that transferring RNAi structure of 3-terminal part of cp gene into potato is one of the better PVX-resistant transgenic strategies.

关 键 词：马铃薯X病毒 外壳蛋白 基因克隆 核苷酸序列分析 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]