双T-DNA表达载体转化大豆的研究  

AGROBACTERIUM-MEDIATED TRANSFORATION OF SOYBEAN WITH THE EXPRESSION VECTOR CARRYING TWO SEPARATE T-DNAs

作　　者：张秀春[1] 郭丽琼[1] 吴坤鑫[1] 林俊扬[2] 林俊芳[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所热带生物技术国家重点实验室,海口571101 [2]福建省莆田市农业科学研究所,莆田351100

出　　处：《大豆科学》2005年第4期291-295,共5页Soybean Science

基　　金：国家自然科学基金项目(30371000);广东省自然科学基金项目(032239);广东省科技攻关项目(2005B210201009);华南农业大学校长基金项目(5100-K03003)

摘　　要：利用含有筛选标记基因和目的基因Δ6-脂肪酸脱氢酶基因的双T-DNA共转化表达载体pLIN61,经农杆菌EHA101介导,采用子叶节转化法转化大豆,使用除草剂Glufosinate作为筛选剂,获得了一批携带有玻璃苣Δ6-脂肪酸脱氢酶基因的转基因大豆,转化效率为2.0%,共转化频率及实际转化率分别为56.25%、1.12%。PCR检测和Southern杂交结果证明,玻璃苣D6D已经整合到大豆的基因组上。RT-PCR检测结果显示,玻璃苣Δ6-脂肪酸脱氢酶基因在转基因大豆的转录水平上得到了表达。The co-transformation expression vector, pLIN61 carrying two separate T-DNAs with selectable marker gene and △^6-desaturase gene was used to transformed soybean cotyledonary nodes mediated by Agrobacteriurn turnefaciens strain EHA101. A batch of transgenic soybeans harboring △^6-desaturase gene were obtained through Glufosinate screening. Research results demonstrate that the transformation efficiency, co-transformation frequency and real transformation frequency wrer 2.0%, 56.25% and 1.12% respectively. PCR detection and Southern analysis confirm that borage △^6-desaturase gene Gas integrated into soybean genome. RT-PCR detection showed that borage △^6-desaturase gene had expressed at transcriptional level in the transgenic soybeans.

关 键 词：共转化表达载体 △^6-脂肪酸脱氢酶基因 转基因大豆 

分 类 号：S565.1[农业科学—作物学]