Tyurin A A, Kabardaeva K V, Mustafaev O N, Pavlenko O S, Sadovskaya N S, Fadeev V S, Zvonova E A, Goldenkova-Pavlova I V
Institute of Plant Physiology, Russian Academy of Sciences, Moscow, 127276, Russia.
Biochemistry (Mosc). 2018 Mar;83(3):259-269. doi: 10.1134/S0006297918030069.
A recombinant DNA in which the interferon αA (IFN-αA) gene sequence is integrated into a loop region of the gene coding thermostable lichenase was constructed. This approach of insertion fusion with thermostable lichenase is advantageous in terms of increasing the solubility, stability, and production of the fusion partner in soluble form in general and in the periplasm of bacterial cells in particular. Thus, the insertion of IFN-αA into the loop (53 a.a.) of thermostable lichenase from Clostridium thermocellum resulted in effective expression of the soluble form of the recombinant protein in the periplasm of Escherichia coli without any compromise in biological activity of IFN-αA, while the thermostable lichenase retained its ability for functional folding without dramatic loss of its basic activity and thermostability.
构建了一种重组DNA，其中干扰素αA（IFN-αA）基因序列整合到编码热稳定地衣酶的基因的环区。一般而言，尤其是在细菌细胞周质中，这种与热稳定地衣酶进行插入融合的方法有利于提高融合伴侣的溶解性、稳定性以及以可溶形式的产量。因此，将IFN-αA插入来自嗜热栖热放线菌的热稳定地衣酶的环区（53个氨基酸），可在大肠杆菌周质中有效表达重组蛋白的可溶形式，且IFN-αA的生物学活性不受任何影响，同时热稳定地衣酶保留了其功能折叠能力，其基本活性和热稳定性也没有显著丧失。
