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Escherichia coli plasmid vectors containing synthetic translational initiation sequences and ribosome binding sites fused with the lacZ gene.
Gene 1982; 19(2):211-9GENE

Abstract

The construction of a series of Escherichia coli plasmid vectors suitable for assaying the effects of gene control signals fused with the E. coli lacZ gene is reported. A synthetic deoxyoligonucleotide dodecamer 5'-CATGAATTCATG GTACTTAAGTAC-5' containing two translation initiation codons (ATG) separated by an EcoRI site was ligated with a lacZ gene derivative which lacks the codons for the first eight amino acids in plasmid pMC1403 (Casadaban et al., 1980). Two ribosome-binding sequences were synthesised and inserted into the EcoRI site before an ATG, and the effects of these sequences on lacZ gene expression in vivo measured by assaying beta-galactosidase activity. The E. coli ribosomal RNA gene (rrnB) promoter, the tetracycline resistance gene promoter, and a lambda phage promoter were cloned using these plasmids. The plasmids are 9.9 kb in size, have ampicillin resistance as a selectable marker and are generally useful for the detection and in vivo assay of gene control regions.

Authors

No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

6293930

Citation

Thomas, D Y., et al. "Escherichia Coli Plasmid Vectors Containing Synthetic Translational Initiation Sequences and Ribosome Binding Sites Fused With the lacZ Gene." Gene, vol. 19, no. 2, 1982, pp. 211-9.
Thomas DY, Dubuc G, Narang S. Escherichia coli plasmid vectors containing synthetic translational initiation sequences and ribosome binding sites fused with the lacZ gene. Gene. 1982;19(2):211-9.
Thomas, D. Y., Dubuc, G., & Narang, S. (1982). Escherichia coli plasmid vectors containing synthetic translational initiation sequences and ribosome binding sites fused with the lacZ gene. Gene, 19(2), pp. 211-9.
Thomas DY, Dubuc G, Narang S. Escherichia Coli Plasmid Vectors Containing Synthetic Translational Initiation Sequences and Ribosome Binding Sites Fused With the lacZ Gene. Gene. 1982;19(2):211-9. PubMed PMID: 6293930.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Escherichia coli plasmid vectors containing synthetic translational initiation sequences and ribosome binding sites fused with the lacZ gene. AU - Thomas,D Y, AU - Dubuc,G, AU - Narang,S, PY - 1982/9/1/pubmed PY - 1982/9/1/medline PY - 1982/9/1/entrez SP - 211 EP - 9 JF - Gene JO - Gene VL - 19 IS - 2 N2 - The construction of a series of Escherichia coli plasmid vectors suitable for assaying the effects of gene control signals fused with the E. coli lacZ gene is reported. A synthetic deoxyoligonucleotide dodecamer 5'-CATGAATTCATG GTACTTAAGTAC-5' containing two translation initiation codons (ATG) separated by an EcoRI site was ligated with a lacZ gene derivative which lacks the codons for the first eight amino acids in plasmid pMC1403 (Casadaban et al., 1980). Two ribosome-binding sequences were synthesised and inserted into the EcoRI site before an ATG, and the effects of these sequences on lacZ gene expression in vivo measured by assaying beta-galactosidase activity. The E. coli ribosomal RNA gene (rrnB) promoter, the tetracycline resistance gene promoter, and a lambda phage promoter were cloned using these plasmids. The plasmids are 9.9 kb in size, have ampicillin resistance as a selectable marker and are generally useful for the detection and in vivo assay of gene control regions. SN - 0378-1119 UR - https://www.unboundmedicine.com/medline/citation/6293930/Escherichia_coli_plasmid_vectors_containing_synthetic_translational_initiation_sequences_and_ribosome_binding_sites_fused_with_the_lacZ_gene_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0378-1119(82)90008-7 DB - PRIME DP - Unbound Medicine ER -