高赖氨酸蛋白质基因和bar基因导入水稻及转基因植株的检测  

Transfer of High Lysine-rich Gene and bar Gene into Rice by Microprojectile Bombardment and Detection of Their Transgenic Plants

作　　者：王为民[1] 赵倩[1] 朱登云[1] 于静娟[1] 敖光明[1] 

机构地区：[1]中国农业大学生物学院农业生物技术国家重点实验室,北京10094

出　　处：《农业生物技术学报》2004年第4期363-368,共6页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目资助(No.2001AA212091和No.2002AA212101)。

摘　　要：构建了1个含高赖氨酸蛋白质基因和bar基因的植物表达载体，用基因枪法将其分别导入籼稻(Oryza sativa ssp．indica)中优早5号幼胚和龙特甫B成熟胚诱导的愈伤组织中，筛选后得到16株可育的再生植株。PCR和Southern blot检测结果表明，外源基因已整合到水稻的基因组中。T1代植株表现出对PPT的抗性。对部分T1代转基因水稻种子进行分析表明，中优早5号种子中蛋白质含量(12．07％～16．2％)和赖氨酸含量(0．52％-0．67％)分别提高了35．18％和45．09％，龙特甫B种子中蛋白质含量(11．92％，12．87％)和赖氨酸含量(0．51％-0．57％)分别提高了27．69％和16．15％。The expression vector containing cDNA encoding a lysine-rich protein and bar gene was constructed. The mature embryonic calluses of Longtefu B and the primary embryo of Zhongyouzao 5 were transformed with expression vector by microprojectile bombardment. And sixteen fertile regenerated plants were obtained. Genomic PCR and Southern blotting analysis showed that the foreign genes were integrated into the rice genome. Resistance analysis showed that some T1 plants were resistant to herbicide. The analysis of protein and lysine contents revealed that in Zhongyouzao 5 transgenic seeds, the protein content (12.07%~16.20%) increased 35.18% as well as the lysine content (0.52%~0.67%) increased 45.09%, and in Longtefu B transgenic seeds the protein content (11.92%~12.87%) increased 27.69% as well as the lysine content (0.51%~0.57%) increased 16.15%.

关 键 词：高赖氨酸 转基因植株 基因导入 Southern 中优早5号 赖氨酸含量 蛋白质含量 植物表达载体 BAR基因 基因枪法 愈伤组织 再生植株 检测结果 blot 外源基因 植株表现 水稻种子 成熟胚 sp. PCR 基因组 PPT 分析表 籼稻 幼胚 

分 类 号：S513[农业科学—作物学] S511