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用缺口双链DNA的定向突变方法分别将胰岛素前体中B链第 2 2、2 8、2 9和 3 0位改变为Asp、Lys、Pro和Lys,酵母分泌表达的前体经胰蛋白酶直接酶切 ,得到重组 [B2 2Asp、B2 8Lys、B2 9Pro、B3 0Lys]人胰岛素。它与受体的结合能力约为猪胰岛素的 6% ,而体内生物活力保留 5 0 %。通过FPLC分子筛测定其自身结合能力 ,在生理条件下浓度达 10 -4mol/L时它以单体形式存在。作为可抗胰蛋白酶酶解的单体胰岛素类似物 ,它可能具有一定的应用前景
Using the mutagenesis method of double-stranded DNA, the 2, 22, 8, 29 and 30 of the B chain in the insulin precursor were changed to Asp, Lys, Pro and Lys respectively, and the precursor secreted by the yeast was directly trypsinized Restriction enzyme digestion gave recombinant [B2 2Asp, B2 8Lys, B2 9Pro, B3 0Lys] human insulin. Its binding capacity with receptors is about 6% of porcine insulin, while the biological activity in vivo remains 50%. Its self-binding capacity was measured by FPLC molecular sieve, which exists as a monomer when the concentration is 10 -4 mol / L under physiological conditions. As a monomeric insulin analog that is antitrypsin-soluble, it may have some promise