有性杂交获得同时表达两种外源基因的转基因胡萝卜的研究

Study of the sexual hybridization method for transgenic carrot with two target genes

以胡萝卜下胚轴为外植体,通过分别携带2种不同外源目的基因片段的LBA4404根癌农杆菌介导转化,经PCR,Southern-blot和RT-PCR分析鉴定,选取各自转化成功并正常转录的T1代植株进行有性杂交,获得T1代杂交种子.同年秋季播种长出T2代植株,经PCR、Western blot检测及Elisa分析表明,得到同时含有2种目的基因且2种目的蛋白表达量都较高的转基因胡萝卜.

Hypocotyls of carrot （Daucus carota var. sativa） were infected with Agrobacterium tumefaciens LBA4404 carrying the target gene. After kanamycin-resistant selection for several times,the regenerated carrot plantlets were analyzed further by PCR,Southern blot and RT-PCR. The results confirmed that the target gene had been introduced into the genome and could be transcripted regularly. The carrot plants （T2generation） integrated different target gene were obtained by sexual hybridization between two groups of transgenic carrots which contained one kind of target gene respectively. The results of PeR,Western blot and Elisa of T2 transgenic plants showed that the two target genes were expressed in roots of transgenie carrots.