15 μg (lyophilized plasmid contains salt of TE buffer)
Storage:
Store at -20ºC
Plasmid copy number:
Medium copy
Replicon:
pMB1
Antibiotic resistance:
Kanamycin
Feature and recommendation:
Stringent regulation with lac repressor
Description:
pETIK is a medium copy number, kanamycin resistant, stringent controllable T7 bacterial expression vector. The T7 expression system is one of the strongest expression systems and has been widely used with a coupling of BL21 (DE3) E. coli cell. T7 RNA polymerase gene is integrated in a genome of BL21(DE3) under control of lacUV5 promoter. Upon addition of isopropyl-1-thio-β-D-galactopyranoside (IPTG), T7 RNA polymerase is expressed in the BL21(DE3) cells harboring pETIK vector, and it induces a high-level protein expression from T7 promoter of pETIK. The pETIK has a lacI gene, which represses T7 RNA polymerase gene in the absence of IPTG. The regulation with lac repressor is beneficial to repress a basal level protein expression and to maintain a recombinant plasmid in BL21 (DE3) cell.
Plasmid Map:
Reconstitution: Resuspend the lyophilized pETIK with 15 µl of sterile water to make 1 μg/µl plasmid in 1 × TE buffer. After reconstitution, store at - 20ºC.
Reference:
1) Studier, F.W. and Moffatt, B.A., J. Mol. Biol. 189 (1986) 113–130.
2) Moffatt, B.A. and Studier, F.W., Cell 49 (1987) 221-227
3) Pan, S. and Malcom, B.A., BioTechniques 29 (2000), 1234–1238
General reference in this Product Information
Sambrook, J. and Russell, D.W. (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring
Harbor Laboratory Press, Cold Spring Harbor, NY.