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A broad-host-range vector system for cloning and translational lacZ fusion analysis.
Plasmid 1988; 19(3):175-88P

Abstract

A broad-host-range vector system for studying translational fusions was constructed. The region that retains the origin of replication, nic, mob, and rep genes of the broad-host-range plasmid RSF1010 was isolated as either an HincII or a PstI-PvuII restriction fragment. These restriction fragments were ligated to tetracycline, kanamycin, or streptomycin/spectinomycin resistance genes to generate plasmids pUI501, pUI511, pUI504, and pUI506. A functional lacZ gene lacking downstream lac operon sequences together with the lac promoter was constructed from plasmids pMC1871 and pUC18. This lacZ gene was inserted into pUI501 and pUI511 to generate plasmids pUI502, pUI503, pUI512, and pUI513. An oligodeoxynucleotide sequence that carries three unique blunt-end restriction sites was synthesized, annealed, and ligated in frame to the amino-terminal end of the lacZ gene in each of these plasmids. This multiple cloning sequence will allow translational fusions to the lacZ gene in all three reading frames. The stability of these plasmids and the expression of the lacZ gene in both Escherichia coli and Rhodobacter sphaeroides were studied.

Authors+Show Affiliations

University of Illinois, Department of Microbiology, Urbana 61801.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

2852814

Citation

Tai, T N., et al. "A Broad-host-range Vector System for Cloning and Translational lacZ Fusion Analysis." Plasmid, vol. 19, no. 3, 1988, pp. 175-88.
Tai TN, Havelka WA, Kaplan S. A broad-host-range vector system for cloning and translational lacZ fusion analysis. Plasmid. 1988;19(3):175-88.
Tai, T. N., Havelka, W. A., & Kaplan, S. (1988). A broad-host-range vector system for cloning and translational lacZ fusion analysis. Plasmid, 19(3), pp. 175-88.
Tai TN, Havelka WA, Kaplan S. A Broad-host-range Vector System for Cloning and Translational lacZ Fusion Analysis. Plasmid. 1988;19(3):175-88. PubMed PMID: 2852814.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A broad-host-range vector system for cloning and translational lacZ fusion analysis. AU - Tai,T N, AU - Havelka,W A, AU - Kaplan,S, PY - 1988/5/1/pubmed PY - 1988/5/1/medline PY - 1988/5/1/entrez SP - 175 EP - 88 JF - Plasmid JO - Plasmid VL - 19 IS - 3 N2 - A broad-host-range vector system for studying translational fusions was constructed. The region that retains the origin of replication, nic, mob, and rep genes of the broad-host-range plasmid RSF1010 was isolated as either an HincII or a PstI-PvuII restriction fragment. These restriction fragments were ligated to tetracycline, kanamycin, or streptomycin/spectinomycin resistance genes to generate plasmids pUI501, pUI511, pUI504, and pUI506. A functional lacZ gene lacking downstream lac operon sequences together with the lac promoter was constructed from plasmids pMC1871 and pUC18. This lacZ gene was inserted into pUI501 and pUI511 to generate plasmids pUI502, pUI503, pUI512, and pUI513. An oligodeoxynucleotide sequence that carries three unique blunt-end restriction sites was synthesized, annealed, and ligated in frame to the amino-terminal end of the lacZ gene in each of these plasmids. This multiple cloning sequence will allow translational fusions to the lacZ gene in all three reading frames. The stability of these plasmids and the expression of the lacZ gene in both Escherichia coli and Rhodobacter sphaeroides were studied. SN - 0147-619X UR - https://www.unboundmedicine.com/medline/citation/2852814/A_broad_host_range_vector_system_for_cloning_and_translational_lacZ_fusion_analysis_ L2 - https://medlineplus.gov/genesandgenetherapy.html DB - PRIME DP - Unbound Medicine ER -