以农杆菌为介导花生的遗传转化研究 I.质粒载体的分子鉴定及农杆菌菌株的转换

Genetic Transformation of Peanut (Arachis hypogaea L.) Mediated by Agrobacterium tumefaciens I. Molecular Identification of Plasmid Vector and Substitute of A. tumefaciens Strains

以农杆菌为介导将外源基因转入花生是花生遗传转化的主要途径之一,本研究利用含几丁质酶和β 1,3 葡聚糖酶基因的植物双价表达载体pCGⅡ进行花生的遗传转化研究。首先对质粒表达载体进行分子鉴定,对LBA4404菌株进行抗生素抗性试验,并针对原有菌株(HLBA4404)对抗生素的不敏感性将其换为另一菌株(NLBA4404),为花生的遗传转化奠定物质基础。

Agrobacterium tumefaciensmediated gene transfer was the main way to produce transgenic plants in peanut. To conduct transgenic research on peanut using binary vectors pCGⅡharboring Chitinase and Glucanase genes, molecular identification of plasmid expression vector was carried out and the antibiotic resistance of LBA4404 strain was tested before genetic transformation. Considering the insensitive characteristics to antibiotics of the old Agrobacterium tumefaciens strain, a new strain was adopted, which provided new genetic transformation system for extensive peanut transgenic research.