A tightly regulated inducible expression system utilizing the fim inversion recombination switch

被引:55
作者
Ham, TS
Lee, SK
Keasling, JD [1 ]
Arkin, AP
机构
[1] Univ Calif Berkeley, Dept Bioengn, Berkeley, CA 94720 USA
[2] Univ Calif Berkeley, Dept Chem Engn, Berkeley, CA 94720 USA
[3] Lawrence Berkeley Lab, Phys Biosci Div, Berkeley, CA 94720 USA
关键词
araBAD promoter; E; coli; expression vector; fim inversion; genetic switch; GFIP expression;
D O I
10.1002/bit.20916
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 fim inversion system of Escherichia coli (E. colt) can behave as a unidirectional switch in an efficient manner. We have developed a new expression system for E. coli, comprising the arabinose-inclucible fimE gene and the fim invertible DNA segment containing a constitutively active promoter. In this system, the target gene is cloned with the promoter in the OFF orientation, resulting in no transcribed product. When induced by arabinose, the active promoter is switched to the ON orientation via FimE-catalyzed DNA inversion, and the gene is expressed. Our expression system exhibited very tightly controlled basal expression and high induced expression, with simple induction by inexpensive arabinose. These characteristics make our system suitable for large-scale expression or for production of toxic proteins. (c) 2006 Wiley Periodicals, Inc.
引用
收藏
页码:1 / 4
页数:4
相关论文
共 14 条
[1]   Green fluorescent protein as a real time quantitative reporter of heterologous protein production [J].
Albano, CR ;
Randers-Eichhorn, L ;
Bentley, WE ;
Rao, G .
BIOTECHNOLOGY PROGRESS, 1998, 14 (02) :351-354
[2]   TIGHTLY REGULATED TAC PROMOTER VECTORS USEFUL FOR THE EXPRESSION OF UNFUSED AND FUSED PROTEINS IN ESCHERICHIA-COLI [J].
AMANN, E ;
OCHS, B ;
ABEL, KJ .
GENE, 1988, 69 (02) :301-315
[3]   QUANTITATIVE-ANALYSIS OF RIBOSOME BINDING-SITES IN ESCHERICHIA-COLI [J].
BARRICK, D ;
VILLANUEBA, K ;
CHILDS, J ;
KALIL, R ;
SCHNEIDER, TD ;
LAWRENCE, CE ;
GOLD, L ;
STORMO, GD .
NUCLEIC ACIDS RESEARCH, 1994, 22 (07) :1287-1295
[4]   TYPE-1 FIMBRIATION AND FIME MUTANTS OF ESCHERICHIA-COLI K-12 [J].
BLOMFIELD, IC ;
MCCLAIN, MS ;
PRINC, JA ;
CALIE, PJ ;
EISENSTEIN, BI .
JOURNAL OF BACTERIOLOGY, 1991, 173 (17) :5298-5307
[5]   The regulation of pap and type 1 fimbriation in Escherichia coli [J].
Blomfield, IC .
ADVANCES IN MICROBIAL PHYSIOLOGY, VOL 45, 2001, 45 :1-49
[6]   TRANSLATIONAL INITIATION ON STRUCTURED MESSENGERS - ANOTHER ROLE FOR THE SHINE-DALGARNO INTERACTION [J].
DESMIT, MH ;
VANDUIN, J .
JOURNAL OF MOLECULAR BIOLOGY, 1994, 235 (01) :173-184
[7]   TIGHT REGULATION, MODULATION, AND HIGH-LEVEL EXPRESSION BY VECTORS CONTAINING THE ARABINOSE P-BAD PROMOTER [J].
GUZMAN, LM ;
BELIN, D ;
CARSON, MJ ;
BECKWITH, J .
JOURNAL OF BACTERIOLOGY, 1995, 177 (14) :4121-4130
[9]   A propionate-inducible expression system for enteric bacteria [J].
Lee, SK ;
Keasling, JD .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2005, 71 (11) :6856-6862
[10]   Improved gfp and inaZ broad-host-range promoter-probe vectors [J].
Miller, WG ;
Leveau, JHJ ;
Lindow, SE .
MOLECULAR PLANT-MICROBE INTERACTIONS, 2000, 13 (11) :1243-1250