Wolber V, Maeda K, Schumann R, Brandmeier B, Wiesmüller L, Wittinghofer A
Max-Planck-Institut für medizinische Forschung, Abteilung Biophysik, Heidelberg, FRG.
Biotechnology (N Y). 1992 Aug;10(8):900-4. doi: 10.1038/nbt0892-900.
We have constructed a series of Escherichia coli expression vectors that produce high yields of fusion proteins containing the C-terminal fragment of light meromyosin (LMM) from rabbit fast skeletal muscle. The fusion proteins retain the ability of LMM to form polymers in low salt and to be soluble in high salt. Thus they can be easily purified from bacterial extracts with a high salt-low salt extraction procedure and still retain their biochemical properties. We demonstrate the utility of this system for the heterologous production and simple purification of LMM fusions of p21H-ras, the neurofibromatosis type I protein and the Tat and protease proteins of HIV-1.
我们构建了一系列大肠杆菌表达载体，这些载体能够高产融合蛋白，该融合蛋白包含来自兔快肌骨骼肌的轻酶解肌球蛋白（LMM）的C末端片段。这些融合蛋白保留了LMM在低盐条件下形成聚合物以及在高盐条件下可溶的能力。因此，它们可以通过高盐-低盐提取程序从细菌提取物中轻松纯化出来，并且仍然保留其生化特性。我们展示了该系统在异源生产和简单纯化p21H-ras、I型神经纤维瘤蛋白以及HIV-1的Tat和蛋白酶蛋白的LMM融合蛋白方面的实用性。
