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A totally synthetic plasmid for general cloning, gene expression and mutagenesis in Escherichia coli.
Gene 1990; 94(1):103-7GENE

Abstract

A first totally synthetic Escherichia coli plasmid has been designed, constructed and shown to be a functional, stable, high-copy cloning vector. The FokI method of gene synthesis [Mandecki and Bolling, Gene 68 (1988) 101-107] was used to assemble the plasmid from 30 oligodeoxyribonucleotides. The plasmid contains synthetic modules for the beta-lactamase-encoding gene (bla), replication origin, lacZ gene fragment and multicloning site. The plasmid is patterned after the pUC-type plasmids and has a copy number similar to that of pUC plasmids. The major changes introduced include the removal of nearly 50% of the restriction sites present in pUC plasmids, reduction of plasmid size to 2050 bp, and introduction of transcription terminators downstream from both the bla gene and lacZ fragment. The changes facilitate a number of techniques, such as cloning, mutagenesis, expression and restriction analysis.

Authors+Show Affiliations

Corporate Molecular Biology, Abbott Laboratories, Abbott Park, IL 60064.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

2227445

Citation

Mandecki, W, et al. "A Totally Synthetic Plasmid for General Cloning, Gene Expression and Mutagenesis in Escherichia Coli." Gene, vol. 94, no. 1, 1990, pp. 103-7.
Mandecki W, Hayden MA, Shallcross MA, et al. A totally synthetic plasmid for general cloning, gene expression and mutagenesis in Escherichia coli. Gene. 1990;94(1):103-7.
Mandecki, W., Hayden, M. A., Shallcross, M. A., & Stotland, E. (1990). A totally synthetic plasmid for general cloning, gene expression and mutagenesis in Escherichia coli. Gene, 94(1), pp. 103-7.
Mandecki W, et al. A Totally Synthetic Plasmid for General Cloning, Gene Expression and Mutagenesis in Escherichia Coli. Gene. 1990 Sep 28;94(1):103-7. PubMed PMID: 2227445.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A totally synthetic plasmid for general cloning, gene expression and mutagenesis in Escherichia coli. AU - Mandecki,W, AU - Hayden,M A, AU - Shallcross,M A, AU - Stotland,E, PY - 1990/9/28/pubmed PY - 1990/9/28/medline PY - 1990/9/28/entrez SP - 103 EP - 7 JF - Gene JO - Gene VL - 94 IS - 1 N2 - A first totally synthetic Escherichia coli plasmid has been designed, constructed and shown to be a functional, stable, high-copy cloning vector. The FokI method of gene synthesis [Mandecki and Bolling, Gene 68 (1988) 101-107] was used to assemble the plasmid from 30 oligodeoxyribonucleotides. The plasmid contains synthetic modules for the beta-lactamase-encoding gene (bla), replication origin, lacZ gene fragment and multicloning site. The plasmid is patterned after the pUC-type plasmids and has a copy number similar to that of pUC plasmids. The major changes introduced include the removal of nearly 50% of the restriction sites present in pUC plasmids, reduction of plasmid size to 2050 bp, and introduction of transcription terminators downstream from both the bla gene and lacZ fragment. The changes facilitate a number of techniques, such as cloning, mutagenesis, expression and restriction analysis. SN - 0378-1119 UR - https://www.unboundmedicine.com/medline/citation/2227445/A_totally_synthetic_plasmid_for_general_cloning_gene_expression_and_mutagenesis_in_Escherichia_coli_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0378-1119(90)90474-6 DB - PRIME DP - Unbound Medicine ER -