RRC ID 33353
Author Dersch P, Fsihi H, Bremer E.
Title Low-copy-number T7 vectors for selective gene expression and efficient protein overproduction in Escherichia coli.
Journal FEMS Microbiol Lett
Abstract A set of low-copy-number vectors (pPD) has been constructed that permit selective gene expression and high-level protein overproduction in Escherichia coli, based on the bacteriophage T7 RNA polymerase/T7 promoter system. These plasmids carry a chloramphenicol resistance gene (cat) as a selective marker and an extended multiple cloning site for convenient gene cloning. Their replication is mediated by ori sequences derived from the low-copy-number vector pSC101. The efficient T7 gene 10 promoter present on these vectors allows selective and high-level transcription of cloned genes carrying their own translational initiation signals. In addition, low-copy-number T7 vectors were constructed that permit expression of genes lacking their own transcription and translation initiation elements by providing a ribosome binding site, an ATG start codon and a multiple cloning site devised for the cloning in all three reading frames. The pPD expression vectors were used to achieve high-level overproduction of the E. coli integral outer membrane protein Tsx, and the cytoplasmic enzymes beta-galactosidase (beta Gal) and UTP:alpha-D-glucose-1-phosphate uridylyltransferase (GalU). The characteristics of these low-copy-number T7 expression vectors should prove very useful for the cloning and high-level overexpression of genes whose gene products are deleterious to the E. coli host.
Volume 123(1-2)
Pages 19-26
Published 1994-10-15
DOI 10.1111/j.1574-6968.1994.tb07195.x
PMID 7988888
MeSH Bacterial Outer Membrane Proteins / biosynthesis Bacterial Outer Membrane Proteins / genetics Bacterial Proteins / biosynthesis* Bacterial Proteins / genetics Bacteriophage T7 / genetics* Base Sequence Escherichia coli / genetics* Escherichia coli / metabolism Escherichia coli Proteins* Gene Expression Regulation, Bacterial Molecular Sequence Data Plasmids / genetics Receptors, Virus* beta-Galactosidase / biosynthesis beta-Galactosidase / genetics
IF 1.987
Times Cited 30
WOS Category MICROBIOLOGY
Resource
Prokaryotes E. coli pPD1 pPD100 pPD101 pPD110 pPD111 pPD112