谷胱甘肽还原酶基因的表达载体构建及对马铃薯的转化  

The Construction of the Gluthione Reductase Gene and Potato Gene Transformation

作　　者：林久生[1] 王根轩[1] 梅 

机构地区：[1]兰州大学

出　　处：《中国农业科学》2001年第2期223-226,T001,共5页Scientia Agricultura Sinica

基　　金：国家自然科学基金(39770447); 国家重点基础研究专项经费(G199990111705)资助项目

摘　　要：利用 ｐＧＲ２０２和 ｐＢｉｎ１９两种质粒经过 ｐＢＩ ２２２的改建，含有 ＣａＭＶ３５Ｓ启动子和 Ｎｏｓ终止子的 ＣＮ区ＤＮＡ片段与 ｐＢｉｎ１９的 Ｂｉｎ区 ＤＮＡ片段的不对称连接和目的基因 ＧＲ的 ｃＤＮＡ重组于真核表达双元载体三步亚克隆，筛选出一正向插入的谷胱甘肽还原酶基因的真核表达双元载体质粒ｐＢｉｎ－ＣＮ－ＧＲ。再利用此质粒转化的农杆菌ＬＢＡ４４０４进行马铃薯的遗传转化．并分化出卡那霉素抗性苗。过氧化物同工酶和酯酶同工酶谱带显示转化材料与对照之间存在显著的差异．表明转化材料的基因表达发生了变化。对转化影响因素的研究发现培养基的组成成分、选择压（Ｋｍ）添加浓度及加入时机是影响转化率的重要因素。Three plasmids (pB I 222, pGR202 and pBin19) were used to construct expression binary vector of the gene of glutathione reductase. Thereafter, the constructed plasmid was introduced into Agrobactium tumefaciens LBA4404. The transformation of potato ( Solanum tuberosum ) by LBA4404 containing the gene of GR was performed. POD and esterase isoenzyme patterns had significant differences between the transformed material and the control, which indicated some changes in gene expression. Different factors influencing transformation included the components of the medium, the concentration of selective antibiotics and time interval before selection were discussed.

关 键 词：马铃薯 根癌农杆菌 表达 基因转化 GR 谷腕甘肽还原酶 

分 类 号：S532[农业科学—作物学]