NEW APPROACHES TO INCREASE THE EXPRESSION AND STABILITY OF CLONED FOREIGN GENES IN ESCHERICHIA-COLI

被引:8
作者
LUKACSOVICH, T [1 ]
BALIKO, G [1 ]
OROSZ, A [1 ]
BALLA, E [1 ]
VENETIANER, P [1 ]
机构
[1] CHEM WORKS GEDEON RICHTER LTD,H-1475 BUDAPEST 10,HUNGARY
关键词
Consensus; P[!sub]2[!/sub] Promoter; Replacement; rrnB gene; Terminator;
D O I
10.1016/0168-1656(90)90072-J
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
A family of expression plasmid vectors were constructed by fusing the strong P2 promoter of the rrnB gene of Escherichia coli (coding for ribosomal RNA) to the lac operator, thereby eliminating regulatory sequences from the rrnB gene and placing the expression under lac repressor control. This promoter proved to be stronger in vivo than the well-known consensus tac promoter, and its strength could be further increased by converting the sequence to consensus. The stability of the recombinant proteins could be increased by fusion to various lengths of the N-terminal end of β-galactosidase, or by inserting a synthetic oligonucleotide, coding for heptathreonine. A new method was developed for the stabilization of recombinant plasmids without antibiotic selection, based on the presence of an essential gene on the plasmid and its absence from the chromosome. The application of this method is illustrated by the example of a plasmid expressing human proinsulin. © 1993.
引用
收藏
页码:243 / 250
页数:8
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