人血小板衍生生长因子BB的原核高表达及其包涵体复性

High-Level Expression of Recombinant Human PGDF-BB in Escherichia coli and its Renaturation

目的:在原核系统内获得高表达的人血小板衍生生长因子BB(PGDF-BB),并对形成的包涵体进行复性。方法:对PGDF-BB核酸编码序列进行优化,构建pET-22b-PGDF-BB表达载体,以提高PCDF-BB的表达量;优化PGDF-BB包涵体复性条件,提高蛋白复性率和生物活性。结果:构建了pET-22b-PGDF-BB高效表达载体,原核表达的重组人PGDF-BB占细菌总蛋白的25%,PGDF-BB包涵体复性率达到15%。结论:对表达序列的优化设计可显著提高蛋白的表达量,复性方法的改良提高了蛋白的复性率和生物活性。

Objective： To improve the expression level of human platelet-derived growth factor BB（PGDF-BB） in E.coli and increase the renaturation ratio of inclusion body. Methods： The sequence of PGDF-BB designed by ancillary software BioSun was synthesized and inserted into plasmid pET-22b to construct pET-PGDF-BB. The pET- PGDF-BB was transformed into E.coli, and PGDF-BB was expressed by induction of IPTG. Then the condition and procedure of renaturation were optimized to improve renaturation ratio and biological activity. Results： After codon optimization, the expression of PGDF-BB was up to 25 % of total bacterial protein. The renaturation ratio of recombinant PGDF-BB was over 15%. Conclusion： The optimization methods applied in this study contributed to improve PGDF-BB expression level and its renaturation ratio.