Danley D E, Geoghegan K F, Scheld K G, Lee S E, Merson J R, Hawrylik S J, Rickett G A, Ammirati M J, Hobart P M
Pfizer Central Research, Groton, CT 06340.
Biochem Biophys Res Commun. 1989 Dec 29;165(3):1043-50. doi: 10.1016/0006-291x(89)92707-1.
A plasmid vector was used to express the HIV-1 pol open reading frame under the regulation of the bacterial trp promoter in Escherichia coli. This expression system has been used as a source of recombinant viral protease. The self-processed active enzyme was recovered from a soluble fraction of a bacterial cell lysate and purified by a procedure involving four steps of chromatography. The protocol yielded 0.3 mg of protease for each liter of bacterial culture. The protease formed tetragonal bipyramidal crystals which have been used in high-resolution X-ray diffraction studies.
一种质粒载体被用于在大肠杆菌中，在细菌色氨酸启动子的调控下表达HIV-1 pol开放阅读框。该表达系统已被用作重组病毒蛋白酶的来源。自加工的活性酶从细菌细胞裂解物的可溶性部分中回收，并通过包含四个色谱步骤的程序进行纯化。该方案每升细菌培养物可产生0.3毫克蛋白酶。该蛋白酶形成了四方双锥晶体，已用于高分辨率X射线衍射研究。
