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将人生长激素释放因子 ( h GRF)基因和编码 L-天门冬酰胺酶 ( L- ans B) C-末端 12 6个氨基酸残基的基因片断进行融合表达 ,融合蛋白在菌体总蛋白中的比率达到 30 %左右 ,比和 L- ans B全基因融合表达时高得多 ,表达条件优化后 ,比率可增加到 4 5%。融合蛋白以不溶性的包含体形式存在。包含体的尿素抽提液经乙醇分级沉淀后得到较纯的融合蛋白 ,60 mmol/ L的盐酸水解融合蛋白 ,通过 SDS- PAGE和电喷雾质谱分析 ,融合蛋白释放出一分子量为 52 35.59Da的物质 ,而 GRF的分子量是 52 35.4 8Da,也就是说融合蛋白酸水解释放出了 GRF
Fusion of human growth hormone releasing factor (h GRF) gene and gene fragment encoding 12 C-terminal 12 amino acids of L-asparaginase (L-ans B) The rate is about 30%, which is much higher than that of the full-gene fusion with L-ans B. After the expression conditions are optimized, the ratio can be increased to 45%. The fusion protein exists as an insoluble inclusion body. The inclusion body of the urea extract was fractionated by ethanol to obtain pure fusion protein, 60 mmol / L hydrochloric acid hydrolysis fusion protein, by SDS-PAGE and electrospray mass spectrometry analysis, the fusion protein released a molecular weight of 5235.59Da Substance, while the molecular weight of GRF is 52 35.4 8Da, which means that the fusion protein acid releases GRF