Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis - PubMed (original) (raw)
Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis
H Shimotsu et al. Gene. 1986.
Abstract
A single-copy integration vector was used for the in vitro construction of translational fusions to the lacZ gene of Escherichia coli. Insertion of a single copy of the lacZ fusion into the B. subtilis chromosome leads to an easily detected Amy- phenotype. A trpE-lacZ fusion was constructed in which the trp promoter directs hybrid beta-galactosidase (beta Gal) synthesis. The level of beta Gal in a wild-type strain carrying the trpE-lacZ fusion in the chromosome is regulated by exogenous tryptophan, while a 5-methyltryptophan-resistant mutant constitutively synthesizes betaGal. A trpF-lacZ fusion was constructed and used to determine the effect of a frameshift mutation in the trpE gene on expression of the trpF-lacZ fusion. The frameshift mutation in trpE led to a three-fold reduction in the levels of the trpF-lacZ fusion. The levels of the betaGal activity of these integrated lacZ fusions appear to provide a quantitative measure of the expression of B. subtilis genes under single-copy conditions.
Similar articles
- A general method for fusion of the Escherichia coli lacZ gene to chromosomal genes in Bacillus subtilis.
Errington J. Errington J. J Gen Microbiol. 1986 Nov;132(11):2953-66. doi: 10.1099/00221287-132-11-2953. J Gen Microbiol. 1986. PMID: 3114418 - cis-acting sites in the transcript of the Bacillus subtilis trp operon regulate expression of the operon.
Kuroda MI, Henner D, Yanofsky C. Kuroda MI, et al. J Bacteriol. 1988 Jul;170(7):3080-8. doi: 10.1128/jb.170.7.3080-3088.1988. J Bacteriol. 1988. PMID: 3133360 Free PMC article. - A broad-host-range vector system for cloning and translational lacZ fusion analysis.
Tai TN, Havelka WA, Kaplan S. Tai TN, et al. Plasmid. 1988 May;19(3):175-88. doi: 10.1016/0147-619x(88)90037-6. Plasmid. 1988. PMID: 2852814 - Uses of lac fusions for the study of biological problems.
Silhavy TJ, Beckwith JR. Silhavy TJ, et al. Microbiol Rev. 1985 Dec;49(4):398-418. doi: 10.1128/mr.49.4.398-418.1985. Microbiol Rev. 1985. PMID: 3005818 Free PMC article. Review. No abstract available. - Organization and regulation of genes encoding biosynthetic enzymes in Bacillus subtilis.
Zalkin H, Ebbole DJ. Zalkin H, et al. J Biol Chem. 1988 Feb 5;263(4):1595-8. J Biol Chem. 1988. PMID: 3123474 Review. No abstract available.
Cited by
- Catabolite repression of the xyl operon in Bacillus megaterium.
Rygus T, Hillen W. Rygus T, et al. J Bacteriol. 1992 May;174(9):3049-55. doi: 10.1128/jb.174.9.3049-3055.1992. J Bacteriol. 1992. PMID: 1569031 Free PMC article. - Similar organization of the sigB and spoIIA operons encoding alternate sigma factors of Bacillus subtilis RNA polymerase.
Kalman S, Duncan ML, Thomas SM, Price CW. Kalman S, et al. J Bacteriol. 1990 Oct;172(10):5575-85. doi: 10.1128/jb.172.10.5575-5585.1990. J Bacteriol. 1990. PMID: 2170324 Free PMC article. - Effect of chromosome location of Bacillus subtilis forespore genes on their spo gene dependence and transcription by E sigma F: identification of features of good E sigma F-dependent promoters.
Sun D, Fajardo-Cavazos P, Sussman MD, Tovar-Rojo F, Cabrera-Martinez RM, Setlow P. Sun D, et al. J Bacteriol. 1991 Dec;173(24):7867-74. doi: 10.1128/jb.173.24.7867-7874.1991. J Bacteriol. 1991. PMID: 1744043 Free PMC article. - Inducible high-level expression of heterologous genes in Bacillus megaterium using the regulatory elements of the xylose-utilization operon.
Rygus T, Hillen W. Rygus T, et al. Appl Microbiol Biotechnol. 1991 Aug;35(5):594-9. doi: 10.1007/BF00169622. Appl Microbiol Biotechnol. 1991. PMID: 1367576 - An A257V mutation in the bacillus subtilis response regulator Spo0A prevents regulated expression of promoters with low-consensus binding sites.
Seredick SD, Seredick BM, Baker D, Spiegelman GB. Seredick SD, et al. J Bacteriol. 2009 Sep;191(17):5489-98. doi: 10.1128/JB.00590-09. Epub 2009 Jul 6. J Bacteriol. 2009. PMID: 19581368 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical