Guo L H, Wang E B, Zhang X Y, Zhu L H, Yu Y J, Wang Y K, Luo G X, Zhou M Y
Shanghai Institute of Cell Biology, Academia Sinica.
Shi Yan Sheng Wu Xue Bao. 1992 Sep;25(3):283-8.
Human insulin A and B chain genes were designed and synthesized by using a rapid and simple method. The synthesized A and B chain genes were cloned separately. The expression (plasmids) pWR 590-HIA and pWR 590-HIB were constructed, and the two plasmids can direct the synthesis of the approximately 590 amino acid-long truncated beta-galactosidases fused to human insulin A or B chains. The fused A or B chain proteins were isolated from the fermented cells and cleaved with BrCN. The resulting mixtures were sulfonated and the sulfonated A and B chains were purified. Human insulin was obtained by using an A and B chain combination method.
采用一种快速简便的方法设计并合成了人胰岛素A链和B链基因。将合成的A链和B链基因分别进行克隆。构建了表达质粒pWR 590 - HIA和pWR 590 - HIB,这两种质粒能够指导合成与人类胰岛素A链或B链融合的约590个氨基酸长的截短型β - 半乳糖苷酶。从发酵细胞中分离出融合的A链或B链蛋白,并用溴化氰进行切割。对所得混合物进行磺化处理,然后纯化磺化后的A链和B链。通过A链和B链组合法获得了人胰岛素。