Promoter/repressor system of Lactobacillus plantarum phage 0g1e:: characterization of the promoters pR49-pR-pL and overproduction of the Cro-like protein Cng in Escherichia coli

被引:12
作者
Kakikawa, M
Watanabe, N
Funawatashi, T
Oki, M
Yasukawa, H
Taketo, A
Kodaira, K
机构
[1] Toyama Univ, Fac Engn, Mol Biol Grp, Toyama 930, Japan
[2] Fukui Med Sch, Dept Biochem 1, Fukui 91011, Japan
关键词
DNA-binding protein; gel-shift assay; operator; primer extension;
D O I
10.1016/S0378-1119(98)00289-3
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The Lactobacillus plantarum phage ogle (42 259 bp) has two repressor-like genes cng and cpg oriented oppositely, accompanied by three potential promoters pR, pL and pR49, and seven operator-like sequences (GATAC-boxes) (Kodaira et al., 1997). In this study, the ogle putative promoters were introduced into the Escherichia coli promoter-detecting plasmid pKK232-8. In E. coli CK111, pR (pKPR1), pL (pKPL1) and pR49 (pKPR49) exhibited distinct CAT activities. When pKPR1 or pKPL1 was coexistent with a compatible plasmid pACYC184 carrying pR-cng (pA4PRCN1), the CAT activity was decreased significantly. On the other hand, cng directed a protein (Cng) of 10.1 kDa in E. coli under the control of T7 promoter. Gel mobility-shift assays demonstrated that Cng binds specifically to a DNA region containing the GATAC-boxes. In addition, primer extension analyses demonstrated that the two sequences pR and pL act as a promoter in L, plantarum as well as in E. coli. These results suggested that the potential promoters pR and pL probably function for the lytic and lysogenic pathways, respectively, and Cng may act as a repressor presumably through the GATAC-boxes as operators. (C) 1998 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:371 / 379
页数:9
相关论文
共 23 条
[1]   High-resolution structure of an engineered cro monomer shows changes in conformation relative to the native dimer [J].
Albright, RA ;
Mossing, MC ;
Matthews, BW .
BIOCHEMISTRY, 1996, 35 (03) :735-742
[2]   SEQUENCE-ANALYSIS OF THE LACTOCOCCUS-LACTIS TEMPERATE BACTERIOPHAGE-BK5-T AND DEMONSTRATION THAT THE PHAGE DNA HAS COHESIVE ENDS [J].
BOYCE, JD ;
DAVIDSON, BE ;
HILLIER, AJ .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1995, 61 (11) :4089-4098
[3]   PLASMID VECTORS FOR THE SELECTION OF PROMOTERS [J].
BROSIUS, J .
GENE, 1984, 27 (02) :151-160
[4]   COMPARATIVE MOLECULAR-BIOLOGY OF LAMBDOID PHAGES [J].
CAMPBELL, A .
ANNUAL REVIEW OF MICROBIOLOGY, 1994, 48 :193-222
[5]   CONSTRUCTION AND CHARACTERIZATION OF AMPLIFIABLE MULTICOPY DNA CLONING VEHICLES DERIVED FROM P15A CRYPTIC MINIPLASMID [J].
CHANG, ACY ;
COHEN, SN .
JOURNAL OF BACTERIOLOGY, 1978, 134 (03) :1141-1156
[6]  
DAVIDSON BE, 1990, FEMS MICROBIOL LETT, V87, P79, DOI 10.1111/j.1574-6968.1990.tb04880.x
[7]   DNA binding by the coliphage 186 repressor protein CI [J].
Dodd, IB ;
Egan, JB .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (19) :11532-11540
[8]   ANALYSIS OF ESCHERICHIA-COLI PROMOTER SEQUENCES [J].
HARLEY, CB ;
REYNOLDS, RP .
NUCLEIC ACIDS RESEARCH, 1987, 15 (05) :2343-2361
[9]   Cloning and nucleotide sequence of the major capsid proteins of Lactobacillus bacteriophage phi gle [J].
Kakikawa, M ;
Oki, M ;
Tadokoro, H ;
Nakamura, S ;
Taketo, A ;
Kodaira, K .
GENE, 1996, 175 (1-2) :157-165
[10]   Characterization of the genes encoding integrative and excisive functions of Lactobacillus phage ogle: Cloning, sequence analysis, and expression in Escherichia coli [J].
Kakikawa, M ;
Oki, M ;
Watanabe, N ;
Yasukawa, H ;
Masamune, Y ;
Taketo, A ;
Kodaira, KI .
GENE, 1997, 185 (01) :119-125