RRC ID 33410
Author Love CA, Lilley PE, Dixon NE.
Title Stable high-copy-number bacteriophage lambda promoter vectors for overproduction of proteins in Escherichia coli.
Journal Gene
Abstract The construction of new high-copy-number (hcn) lambda-promoter expression vectors is described. All these vectors (1) contain tandem lambda pR and pL promoters upstream of an extensive multiple cloning site (MCS) for insertion of genes, (2) direct expression of the lambda cIts857 gene, enabling their use in any Escherichia coli host strain for thermal induction of gene overexpression, and (3) bear the par locus of plasmid pSC101, ensuring their stable maintenance at hcn in the absence of continuous antibiotic selection. Six of the vectors also contain efficient ribosome-binding sites upstream of unique HpaI or NdeI sites in their MCS regions, and two contain sequences that encode N-terminal poly-His. The performance of these vectors was assessed by using them to overproduce the E. coli HMP flavohaemoprotein and the bacteriophage M13 gene II replicator protein.
Volume 176(1-2)
Pages 49-53
Published 1996-10-17
DOI 10.1016/0378-1119(96)00208-9
PII 0378-1119(96)00208-9
PMID 8918231
MeSH Bacterial Proteins / genetics* Bacteriophage lambda / genetics* Base Sequence DNA, Viral Escherichia coli / metabolism Gene Dosage Gene Expression* Genetic Vectors* Hemeproteins / genetics* Molecular Sequence Data Plasmids Promoter Regions, Genetic*
IF 2.984
Times Cited 74
WOS Category GENETICS & HEREDITY
Resource
Prokaryotes E. coli pCL476 pCL478 pND706 pND707 pPL450 pPL451 pPL452