含Cry1Ab/Ac基因的白僵菌工程菌的构建

Construction of beauveria engineering strain containing Cry1Ab/Ac gene

以来源于苏云金芽孢杆菌的Bt-Cry1Ab/Ac基因为目的基因,以潮霉素Hyg为抗性标记基因,以绿色荧光蛋白基因GFP为报告基因的真菌表达载体,采用农杆菌介导真菌遗传转化(ATMT)法,将Cry1Ab/Ac基因插入白僵菌基因组中,先后用潮霉素B抗性筛选、荧光检测、真菌基因组PCR分子检测法、Bt-Cry1Ab/Ac基因检测试纸条多种方法进行转基因工程菌株鉴定,结果证实了目的基因成功插入白僵菌基因组,为白僵菌杀虫活性的深入研究奠定了基础.

Since then,the Bt-Cry1Ab/Ac gene from Bacillus thuringiensis was used as the target gene.Hygromycin (Hyg)was used as the marker gene and green fluorescent protein gene gfp as the reporter gene.The Cry1Ab/Ac gene was inserted into the genome of Beauveria bassiana by Agrobacterium tumefaciens- mediated fungal genetic transformation(ATMT).Hygromycin B resistance screening, fluorescence detection and fungal genomic PCR detection were performed successively.The results showed that the target gene was successfully inserted into Beauveria bassiana gene.The results showed that the Bt-Cry1Ab/Ac gene was successfully inserted into Beauveria bassiana.It laid a foundation for further study on insecticidal activity of Beauveria bassiana.