Schleif R F, Favreau M A
Biochemistry. 1982 Feb 16;21(4):778-82. doi: 10.1021/bi00533a031.
Hypersynthesis of araC protein from Escherichia coli has been accomplished. The araC gene was cloned on plasmid pBR322, and some of the noncoding DNA preceding the araC gene was removed by exonuclease digestion. Finally, a DNA fragment containing the lac promoter and ribosome binding site was placed in front the araC gene. By these means the level of araC protein was increased about 5000-fold above the levels found in wild-type cells. This level of protein permits straight forward purification of sizeable quantities of araC protein.
已实现了来自大肠杆菌的阿拉伯糖操纵子C(araC)蛋白的超量合成。将araC基因克隆到质粒pBR322上,并且通过核酸外切酶消化去除了araC基因之前的一些非编码DNA。最后,将含有乳糖启动子和核糖体结合位点的DNA片段置于araC基因之前。通过这些方法,araC蛋白的水平比野生型细胞中的水平提高了约5000倍。这种蛋白水平使得能够直接纯化出相当数量的araC蛋白。