Sun Qiao-Yang, Ding Ling-Wen, He Liang-Liang, Sun Yong-Bin, Shao Jun-Li, Luo Ming, Xu Zeng-Fu
State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, Guangdong, China.
Anal Biochem. 2009 Nov 1;394(1):144-6. doi: 10.1016/j.ab.2009.07.023. Epub 2009 Jul 19.
In this paper, we report a useful protocol for cloning toxic protein genes. Use of the SOC medium, which is a glucose-containing rich medium, significantly improved the transformation efficiency of a recombinant plasmid containing a toxic plant subtilase SaSBT1 cDNA. Both glucose and rich nutrients present in the SOC medium prevented the unintended activation of the lac promoter carried on the cloning vector, and led to significantly improved transformation efficiency of recombinant plasmids containing toxic protein genes and an increased rate of transformant growth.
在本文中,我们报道了一种用于克隆毒性蛋白基因的有效方案。使用SOC培养基(一种富含葡萄糖的培养基)可显著提高含有毒性植物枯草杆菌蛋白酶SaSBT1 cDNA的重组质粒的转化效率。SOC培养基中存在的葡萄糖和丰富营养成分可防止克隆载体上携带的lac启动子意外激活,并导致含有毒性蛋白基因的重组质粒的转化效率显著提高,以及转化子生长速率增加。