摘要： 为将改造合成的抗虫基因(Cry1Ac-w)导入玉米基因组,以玉米杂交组合PA×PB的胚性愈伤组织为外植体,用双丙氨磷作为抗性愈伤组织的筛选剂,通过农杆菌介导法获得了82个再生植株。经标记基因(bar)和目的基因(Cry1Ac-w)的PCR检测,其中19株为阳性,即为推断的转 Cry1Ac-w 基因抗虫玉米。 对其中2株(w₁和w₂)进行目的基因单酶切的Southern杂交分析,发现w₁和w₂分别为单位点和双位点插入,这也进一步证实目的基因已经整合入玉米基因组中。用Bt-Cry1Ab/Ac金标免疫试纸条进行蛋白质表达的检测,初步证实目标Cry1Ac-w蛋白在玉米中得到表达。

关键词: 玉米, 农杆菌介导法, 苏云金杆菌(Bt), Cry1Ac-w基因, Southern杂交, 免疫检测

Abstract: The purpose of this study was to introduce the modified Cry1Ac-w gene into the corn.Eighty-two regeneration plants were obtained through Agrobacterium -mediated method using embryo callus of the hybrid combination PA×PB as explants and the bialaphos as screening agent,but nineteen plants were putative transgenic plants,confirmed by PCR method of labelled gene(bar)and target gene(Cry1Ac-w).Southern hybridization test(single restriction enzymes digestion)showed successful integration of the foreign Cry1Ac-w gene into the corn genome and the w₁ and w₂ contained one copy and two copies of target gene,respectively.The expressing of Cry1Ac-w protein was confirmed by Bt-Cry1Ab/Ac immune detection strips.

Key words: Corn, Agrobacterium-mediated method, Bacillus thuringiensis (Bt), Cry1Ac-w gene, Southern blot, Immune detection

中图分类号: 

• S435.132

• Q785