霍乱毒素B亚基对胡萝卜的遗传转化

Genetic transformation of CT-B on carrot

以胡萝卜无菌幼苗的下胚轴为外植体,通过携带CTB基因的根癌农杆菌的介导进行转化,在Kan50mg/L和Carb500mg/L的MS3-1固体培养基上诱导形成胚性愈伤组织,转至Kan100mg/L和Carb100mg/L的MS0固体培养基上形成植株,同时还进行了以经过预处理的下胚轴为外植体进行转化的试验,发现低温处理4d的下胚轴的抗性愈伤诱导率较高。经PCR检测、PCR-Southern杂交检测以后,表明CTB基因已整合进胡萝卜的基因组中。RT-PCR检测初步证明了CTB基因的转录。

The hypocotyl of carrot were infected with Agrobacterium tumefaciens containing CTB gene,the explants were transferred to MS31 selection media which contained Kan 50 mg/L and carb 500 mg/L to induce embryogenic callus.Then the callus were transferred to MS0 media to form plantlet,and it was found that the transgenic rate of hypocotyls which were treated of low temperature were higher in all treatments.PCR,PCRSouthern detection of carrot DNA confirmed that the CTB gene had been introduced into the plant genome.RTPCR detection showed primarily the transcription of CTB.