美洲商陆抗真菌蛋白转化玉米的研究和抗病性检测

Maize Transformation of PaAFP Gene and Disease Resistance Assay

研究美洲商陆抗病毒蛋白(PaAFP)基因转入玉米中对玉米纹枯病的抗性。从美洲商陆叶片中获得美洲商陆抗真菌蛋白前体蛋白基因cDNA序列,构建植物表达载体pCAMBIA1300-PaAFP,通过三亲杂交法将其导入根癌农杆菌LBA4404受体菌,转化玉米获得了大量再生转基因植株。PCR、PCR-southern杂交、RT-PCR以及Tris-Tricine-SDS-PAGE检测结果表明,目的基因已经整合到玉米基因组中,并且已经得到转译。

In the paper, plant expression vector pCAMBIA1300-PaAFP was constructed by cloning the PaAFP gene from pokeweed to pCAMBIA-1300. Then, the engineered strain LBA4404 was constructed by conducting the pCAMBIA-1300-PaAFP to A. tumefaciens LBA4404 and was used to transform maize. The PCR, PCR-Southern blotting, RT-PCR, SDS-PAGE analysis indicated that the PaAFP gene had been integrated into the genome of maize and produced the protein.