EXPRESSION OF BIOLOGICALLY-ACTIVE, MATURE HUMAN GRANULOCYTE MACROPHAGE COLONY STIMULATING FACTOR WITH AN ESCHERICHIA-COLI SECRETORY EXPRESSION SYSTEM
EXPRESSION OF BIOLOGICALLY-ACTIVE, MATURE HUMAN GRANULOCYTE MACROPHAGE COLONY STIMULATING FACTOR WITH AN ESCHERICHIA-COLI SECRETORY EXPRESSION SYSTEM
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DOI：
10.1007/bf01570872
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发表时间：
1988-01-01
影响因子：
2.6
通讯作者：
VANKIMMENADE, A
中科院分区：
文献类型：
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作者：
GREENBERG, R;LUNDELL, D;VANKIMMENADE, A
人粒细胞巨噬细胞集落刺激因子（HuGM-CSF）以分泌载体piniiiompa2[10]在大肠杆菌中表达。由此表达的HuGM-CSF蛋白被证明具有忠实的裂解性和生物活性。然而，该蛋白不能通过渗透冲击释放，并且在亚细胞分离时，与外膜组分共同沉积。研究了启动子、载体、宿主菌株、诱导条件和培养基配方对表达水平的影响。其中一些因素在决定大肠杆菌分泌表达系统中HuGM-CSF的最大可达到水平方面起着重要作用。
Human granulocyte-macrophage colony stimulating factor (HuGM-CSF) was expressed periplasmically in Escherichia coli with the secretory vector, pINIIIompA2 [10]. HuGM-CSF protein thus expressed was shown to be faithfully cleaved and biologically active. This protein, however, could not be released by osmotic shock and, on subcellular fractionation, co-sedimented with the outer membrane fraction. The effect of promoters, vectors, host strains, induction conditions and media formulation on expression levels was also evaluated. Some of these factors play a significant role in determining maximal achievable levels of HuGM-CSF in the secretory expression system of E. coli.