RRC ID 33398
Author Mayer MP.
Title A new set of useful cloning and expression vectors derived from pBlueScript.
Journal Gene
Abstract A new set of cloning vectors derived from pBlueScript (Stratagene, La Jolla, CA, USA) is presented. The ampicillin-resistance-encoding gene (ApR) of pBlueScript has been replaced by genes encoding resistance to either kanamycin (KmR) or tetracycline (TcR). The origin of DNA replication (ori), conferring to pBlueScript a very high-copy-number (500-700 copies/chromosome), has been replaced by the pBR322 ori (15-20 copies/chromosome) or the P15A ori (10-12 copies/chromosome) [Sambrook et al.: Molecular Cloning. A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989]. Therefore, eight new vectors with different drug selection markers and low, medium or high plasmid copy-number were created which are compatible with each other (ColE1 ori and P15A ori) and can be selected to replace one another. These vectors were further modified by the insertion of an expression cassette based on the promoter and AraC repressor/activator of the ara operon, which allows high-level expression, extremely tight regulation and very inexpensive induction. High-level expression of one or two genes within the same cell is demonstrated.
Volume 163(1)
Pages 41-6
Published 1995-9-22
PII 037811199500389N
PMID 7557476
MeSH Ampicillin Resistance / genetics* Base Sequence Chromosomes, Bacterial Cloning, Molecular / methods* DNA Primers Gene Expression Genes, Bacterial* Genetic Markers Genetic Vectors* Kanamycin Resistance / genetics* Molecular Sequence Data Mutagenesis, Insertional Operon Plasmids* Polymerase Chain Reaction Replication Origin* Restriction Mapping Tetracycline Resistance / genetics*
IF 2.638
Times Cited 146
Prokaryotes E. coli pMPM-A2 pMPM-A3 pMPM-A4omega pMPM-A5omega pMPM-A6omega pMPM-K1 pMPM-K2 pMPM-K3 pMPM-K4omega pMPM-K5omega pMPM-K6omega pMPM-T1 pMPM-T2 pMPM-T3 pMPM-T4omega pMPM-T5omega pMPM-T6omega