雪莲PBP基因表达载体的构建  

Construction of Plant Expression Vector pXLPBP

作　　者：李萍[1] 艾秀莲[1] 王志方[1] 王博[1] 

机构地区：[1]新疆农科院微生物应用研究所,新疆乌鲁木齐830091

出　　处：《生物技术》2006年第2期11-13,共3页Biotechnology

基　　金：国家自然科学基金项目资助(30160039)

摘　　要：目的:利用新疆雪莲特殊功能基因磷脂酰乙醇胺结合蛋白基因(XLPBP)与基础质粒构建植物表达载体pXLPBP,为介导该基因在植物中表达,以期提高植物抗寒力的转基因研究打下基础。方法:利用设计好的两端加有EcoRⅠ酶切位点的引物,对XLPBP全长基因片段进行PCR扩增,获得700bp左右大小的片段,将其纯化回收并与同样经过EcoRⅠ酶切的质粒pCAMBIA3301连接;然后采用冻融法和电击法,将含XLPBP基因的载体pCAMBIA3301转入根癌农杆菌中。结果:通过一系列分子克隆方法获得含雪莲XLPBP基因的植物表达载体,并经PCR实验证实。结论:利用以自身携带的非编码区为调控序列的XLPBP全长基因和双向表达载体pCAMBIA3301为基础构建植物表达载体,可望提高外源基因表达量。Objectives：Using the XLPBP gene from Saussurea involucrata Kar.et and the plasmid of pCAMBIA3301 to construct plant expression vector pXLPBP. This will be grounded for transformation and expression of the XLPBP gene in plants. Methods： The fragment of barget gene about 700 bp was amplified by PCR, and the site of EcoR Ⅰ was added to both ends of the gene. After purified, it was inserted into expression vector pCAMBIA3301 that was digested by EcoR Ⅰ. Then the recombination was transformed into Agrobacterium twnefecient EHA105 . Results： The plant expression vector pXLPBP was obtained, and conformed sequence by PCR. Conclusion： the vector pXLPBP was constructed by this way, which make it possible to improve expression of the gene.

关 键 词：雪莲 磷脂酰乙醇胺结合蛋白 根癌农杆菌 

分 类 号：Q784[生物学—分子生物学]