Vázquez R, Prieto D, Olóriz M I, de la Riva G A, Selman-Housein G
División de Agricultura, Centro de Ingeniería Genética y Biotecnología, Habana, CUBA.
Rev Latinoam Microbiol. 1995 Jul-Sep;37(3):237-44.
cryIA(b) and cryIA(c) genes encoding active fragments of Bacillus thuringiensis delta-endotoxins were cloned downstream of the pR and pT7 promoters from the lambda and T7 bacteriophages, respectively. cryIA(b) gene was also fused with the gene encoding protein A from Staphylococcus aureus cloned under the control of the pR promoter. There were no remarkable differences in the expression levels of the cloned genes in E. coli, but the Western blot analysis allowed distinct protein quality for the three expression systems. We conclude that the best expression model for the production of delta-endotoxins toxic fragments in E. coli is the one based on lambda pR promoter.
编码苏云金芽孢杆菌δ-内毒素活性片段的cryIA(b)和cryIA(c)基因分别从λ噬菌体和T7噬菌体的pR和pT7启动子下游克隆而来。cryIA(b)基因还与在pR启动子控制下克隆的金黄色葡萄球菌蛋白A编码基因融合。克隆基因在大肠杆菌中的表达水平没有显著差异,但蛋白质印迹分析显示三种表达系统的蛋白质质量不同。我们得出结论,在大肠杆菌中生产δ-内毒素毒性片段的最佳表达模型是基于λ pR启动子的模型。