INTRACELLULAR AND EXTRACELLULAR PRODUCTION OF PROTEINS IN ASPERGILLUS UNDER THE CONTROL OF EXPRESSION SIGNALS OF THE HIGHLY EXPRESSED ASPERGILLUS-NIDULANS GPDA GENE

被引:86
作者
PUNT, PJ
ZEGERS, ND
BUSSCHER, M
POUWELS, PH
VANDENHONDEL, CAMJJ
机构
[1] TNO Medical Biological Laboratory, Rijswijk
关键词
GPD; GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE; PAGE; POLYACRYLAMIDE GEL ELECTROPHORESIS; XGAL; 5-BROMO-4-CHLORO-3-INDOLYL-]BETA-D-GALACTOPYRANOSIDE; XGLUC; 5-BROMO-4-CHLORO-3-INDOLYL-BETA-D-GLUCURONIDE; DESIGNATES PLASMID-CARRIER STATE;
D O I
10.1016/0168-1656(91)90024-P
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The expression in Aspergillus is described of genes, coding for intracellular and extracellular proteins controlled by the promoter region of the constitutively and efficiently expressed glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) of Aspergillus nidulans. Both the homologous gpdA and the heterologous Escherichia coli beta-galactosidase (lacZ) and beta-glucuronidase (uidA) genes could be expressed intracellularly at levels as high as 10-25% of total soluble protein. Efficient extracellular production of A. niger glucoamylase could be achieved with a fusiongene containing the region of the glucoamylase gene coding for the mature protein proceded by a synthetic fungal signal sequence. Extracellular production of a heterologous protein, E. coli beta-glucuronidase, with such a fusion was much less efficient. Only very low levels of beta-glucuronidase were detected in the culture fluid, whereas considerable enzyme activity was detected in the mycelium.
引用
收藏
页码:19 / 33
页数:15
相关论文
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