Bi Q, Zhang J, Huang Y, Su H, Zhou X, Zhu S
College of Life Sciences, Peking University, Beijing, PR China.
Appl Biochem Biotechnol. 2001 Jul;95(1):23-30. doi: 10.1385/abab:95:1:23.
The mutant gene of HV2-K47 was obtained by polymerase chain reaction-directed mutagenesis and expressed in Escherichia coli. Many elements that could affect its expression level were compared. The product was purified to homogeneity via three chromatographic steps--ion exchange, gel filtration, and reverse phase chromatography--on the AKTA Explorer System. The anti-thrombin activity of HV2-K47 is much higher than that of recombinant HV2. Some properties and expression conditions were investigated systematically, which would be useful for further studies of hirudin and other small proteins.
通过聚合酶链反应定向诱变获得HV2-K47突变基因，并在大肠杆菌中表达。比较了许多可能影响其表达水平的因素。通过在AKTA Explorer系统上进行离子交换、凝胶过滤和反相色谱三个色谱步骤，将产物纯化至同质。HV2-K47的抗凝血酶活性远高于重组HV2。系统研究了其一些性质和表达条件，这将有助于水蛭素和其他小蛋白质的进一步研究。
