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Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis.
Gene 1986; 43(1-2):85-94GENE

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.

Authors

No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

3019840

Citation

Shimotsu, H, and D J. Henner. "Construction of a Single-copy Integration Vector and Its Use in Analysis of Regulation of the Trp Operon of Bacillus Subtilis." Gene, vol. 43, no. 1-2, 1986, pp. 85-94.
Shimotsu H, Henner DJ. Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis. Gene. 1986;43(1-2):85-94.
Shimotsu, H., & Henner, D. J. (1986). Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis. Gene, 43(1-2), pp. 85-94.
Shimotsu H, Henner DJ. Construction of a Single-copy Integration Vector and Its Use in Analysis of Regulation of the Trp Operon of Bacillus Subtilis. Gene. 1986;43(1-2):85-94. PubMed PMID: 3019840.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis. AU - Shimotsu,H, AU - Henner,D J, PY - 1986/1/1/pubmed PY - 1986/1/1/medline PY - 1986/1/1/entrez SP - 85 EP - 94 JF - Gene JO - Gene VL - 43 IS - 1-2 N2 - 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. SN - 0378-1119 UR - https://www.unboundmedicine.com/medline/citation/3019840/Construction_of_a_single_copy_integration_vector_and_its_use_in_analysis_of_regulation_of_the_trp_operon_of_Bacillus_subtilis_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0378-1119(86)90011-9 DB - PRIME DP - Unbound Medicine ER -