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2025年8月19日 星期二
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农杆菌介导的蓝色基因转化中国水仙
戴艺民
福建农科院甘蔗研究所
Transformation of Blue Gene in Narcissus tazetta var. chinensis Mediated by Agrobacterium tumefaciens
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摘要 从矮牵牛(Petunia hybrida)花瓣中分离得到编码蓝色基因F3’5’H(类黄酮3’,5’羟基化酶)的Hf2基因(1527bp),从牵牛(Ipomoea nil)花瓣中分离得到dfr基因(二氢黄酮醇4-还原酶,1212bp),将正向Hf2片段和反向dfr片段连接到pBRLys双元载体质粒上,得到具有Ubi(Ubiquitin)组成型启动子的植物表达载体pUbi-Hf2-dfr。利用冻融法将双价重组质粒pUbi-Hf2-dfr导入农杆菌(Agrobacterium tumefaciens)菌株EHA105中,菌落PCR鉴定农杆菌转化子。以中国水仙(Narcissus tazetta var. chinensis)愈伤组织为材料,进行遗传转化。CTAB法提取130株中国水仙遗传转化再生苗的基因组DNA,进行PCR和PCR-Southern blot检测,获得3株阳性转基因植株。
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戴艺民
关键词 蓝色基因中国水仙遗传转化    
Abstract:Hf2 gene(1527bp) encoding flavonoid 3’,5’-hydroxylase (F3’5’H) and dihydroflavonol 4-reductase (dfr) (1212 bp) gene were isolated from Petunia hybrida and Ipomoea nil petals respectively. Hf2 and anti sense dfr genes were subcloned into pBRLys vector together generating pUbi-Hf2-dfr vector which contained Ubiquitin(Ubi) promoter, and then the pUbi-Hf2-dfr was transferred into Agrobacterium tumefaciens strain EHA105. Result of PCR amplification with transformant colony showed that the expression vector pUbi-Hf2-dfr was successfully transferred into Agrobacterium. Hf2 and anti sense dfr genes were transformed into callus of Narcissus tazetta var. chinensis mediated by Agrobacterium. Extracted the genomic DNA of 130 regenerated plants with CTAB method, and then performed PCR with specific primers and electrophoresis results showed that there were 3 regenerated plants with the intergration of Hpt and blue genes. PCR-Southern blotting indicated the same result.
Key wordsblue gene    Narcissus tazetta var. chinensis    genetic transformation
收稿日期: 2009-08-24     
通讯作者: 戴艺民   
引用本文:   
戴艺民. 农杆菌介导的蓝色基因转化中国水仙[J]. , 2010, 18(2): 231-238.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2010/V18/I2/231
 
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