生菜农杆菌遗传转化系统的建立及HIV蛋白基因的导入

Establishment of Genetic Transformation System of Lettuce by Agrobacterium tumefaciens and Introduction of HTV gag-gp120 Gene

通过根癌农杆菌介导,将gag-gp120基因导入到苗龄为5 d的北京生菜无菌苗的子叶片中,同时针对影响农杆菌遗传转化的几个关键因素进行研究试验。结果表明,北京生菜适合的卡那霉素筛选的浓度是7．5 mg／L,添加乙酰丁香酮对农杆菌的浸染影响不大,农杆菌的浸染时间以30 min为宜,农杆菌适宜的浸染浓度为OD600值在0．4～0．8之间,培养时间以3 d效果较好。通过PCR和PCR-Southern分析证明,gag-gp120嵌合基因已经整合到生菜基因组中。

By mediation of agrobacterium tumefaciens, HIV gag-gpl20 gene was transferred into cotyledons of Beijing-lettuces. The transformation influence factors were also studied and regenerated lettuce plants were examined by PCR and PCR-Southern analysis. The results were as follows, the suitable filtration concentration of Kanamycin was 7.5mg/L. The AS additive showed less effects on tumefacien infection, suitable infected time would be 30 minutes, while OD600 of engineering bacterial suspension was 0.4 to 0.8 and 3-days cultivated. It was proved by the PCR and PCR-Southern examination that the target gene had been transferred into lettuce.