无选择标记转高赖氨酸蛋白基因水稻植株的培育  

Cultivation of marker-free transfer of Lysine-rich protein gene on rice

作　　者：段忠卫[1,2] 李希臣[3,4] 张军[1] 刘琦[3,4] 夏善勇[3,4] 刘建新[3,4] 李杰[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030 [2]主要农作物种质创新国家重点实验室,北京100192 [3]黑龙江省农业科学院生物技术研究所,哈尔滨150086 [4]黑龙江省作物与家畜分子育种重点实验室,哈尔滨150086

出　　处：《东北农业大学学报》2013年第7期46-51,共6页Journal of Northeast Agricultural University

基　　金：哈尔滨市青年科技人才项目(RC2009QN002131)

摘　　要：利用农杆菌介导遗传转化方法,将带有分别由Ubi组成型启动子和GluB-4水稻种子特异型启动子调控的GsHLP2/GsHLP8基因、选择标记基因为Bar基因的双T-DNA植物表达载体pTTBUG2、pTTBUG8、pTTBGG2、pTTBGG8转入水稻。共获得转化苗124株,PCR结果表明,高赖氨酸蛋白基因GsHLP2/GsHLP8及Bar基因已经共转化到水稻基因组上,共转化率为61.1%。通过Bialaphos抗性试验对转基因后代进行抗性检测,结果表明T1种子对Bialaphos的抗性出现分离。进而对T1代植株进行PCR检测,获得只含有目的基因而不含有选择标记基因的植株,为培育无选择标记转高赖氨酸蛋白基因水稻提供材料。Using the method of Agrobacterium-mediated genetic transformation, double T-DNA plant expression vector pTTBUG2, pTTBUG8, pTTBGG2, pTTBGG8 which respectively composed of Ubi promoter and GluB-4 rice seed specific promoter regulation of GsHLP21GsHLP8 gene were separatly transfered into rice; and the selective marker of the plant expression vectors were Bar gene. Total 124 transgenic rice had been gained. The result of PCR suggested that Lysine-rich protein gene GsHLP21GsHLP8 and Bar gene had been transformed into the rice genome, the co-transformation rate was 61.1%. The experiment on Bialaphos resistance of the trangenic plant using the PPT as a selective marker showed that the Bialaphos resistance departed in the T1 generation. The PCR detection of the T, plants suggested that the plant which contained only the target gene were obtained and could be used as a marker-free material for cultivation of Lysine-rich protein gene rice.

关 键 词：农杆菌介导 高赖氨酸蛋白 无选择标记 共转化 

分 类 号：S511[农业科学—作物学] Q785[生物学—分子生物学]