番木瓜环斑病毒CP基因同源区段的克隆及植物表达载体的构建  

Cloning of Coat Protein Gene Homologous Segment of Papaya Ringspot Virus and Construction of Its Plant Expression Vectors

作　　者：魏军亚[1] 刘德兵[2] 蔡群芳[1] 黎小瑛[1] 周鹏[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所,海口571101 [2]华南热带农业大学园艺学院,海南儋州571737

出　　处：《热带作物学报》2007年第2期54-59,共6页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金(30560082);国家转基因植物研究与产业化开发专项(JY04-B-02)资助

摘　　要：采用RT-PCR技术对番木瓜环斑病毒CP基因的同源区段进行了克隆和鉴定分析。序列分析结果表明,获得PRSV-CP基因3′端的278bp DNA片段同源率最高。构建了目的基因包含278bp正义链、内含子(PDK内含子)及278bp反义链的RNA介导的植物表达载体p2301-CPU,并通过电激法导入农杆菌中。经PCR及酶切鉴定,证实质粒已被导入获得了农杆菌工程菌株。利用该植物表达载体对番木瓜的遗传转化工作目前正在进行中。The CP gene homologous segment of Papaya ringspot virus （PRSV） was cloned and sequenced using the RT-PCR.The sequence analysis showed that the sequence of DNA segments of 278 bp identified at 3＇terminus of the PRSV-CP-coding region was most homologous. The sense segment and antisense segment of the gene homologous segment of PRSV-CP, and the PDK intron segment were constructed into plant expression vector pCAMBI- A2301 by using a series of cloning methods. The PCR and enzyme digestion showed that the recombinant plasmid was transferred into competent cells of Agrobacterium tumefaciens EHA 105. Transformation of the gene into papaya is still under way.

关 键 词：番木瓜环斑病毒外壳蛋白基因 同源区段 RNA介导植物表达载体 农杆菌工程菌株 

分 类 号：Q78[生物学—分子生物学] S667.903.6[农业科学—果树学]