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PMID: 8054844 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

pJC20 and pJC40--two high-copy-number vectors for T7 RNA polymerase-dependent expression of recombinant genes in Escherichia coli.

Protein expression and purification ·Vol. 5 ·No. 2 ·1994-04-00 ·Pages 133-7

Clos J, Brandau S

Abstract

We report the construction of two plasmid vectors, pJC20 and pJC40, for the expression of recombinant genes in Escherichia coli under the control of T7 RNA polymerase. Their small sizes of ca. 2.4 kb ease the subcloning of large inserts and the high copy numbers obtained result in satisfactory yields in all plasmid preparations. A multiple-cloning site offers sites for directional cloning and nested deletions. In addition, pJC40 encodes a cleavable amino-terminal histidine tail of 10 residues which is added to the gene product, thus allowing purification by metal chelate chromatography. Observed expression yields are in the range of 10% of total bacterial protein for all genes tested in our laboratory.

MeSH Terms
Animals Base Sequence Chromatography, Affinity DNA-Directed RNA Polymerases/genetics Drosophila/genetics Escherichia coli Gene Expression Regulation, Bacterial/drug effects Genes, Synthetic Genetic Vectors Heat-Shock Proteins/biosynthesis,genetics,isolation & purification Humans Isopropyl Thiogalactoside/pharmacology Leishmania major/genetics Molecular Sequence Data Plasmids Promoter Regions, Genetic Recombinant Fusion Proteins/biosynthesis,genetics,isolation & purification Viral Proteins
Chemicals
Heat-Shock Proteins Recombinant Fusion Proteins Viral Proteins Isopropyl Thiogalactoside bacteriophage T7 RNA polymerase DNA-Directed RNA Polymerases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Clos J
Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany.
Brandau S
Article Info
Journal
Protein expression and purification
Abbr.
Protein Expr Purif
ISSN
1046-5928
Published
1994-04-00
Pages
133-7
Language
English
Region
United States
NLM ID
9101496
Subset
IM
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