pGEX-KG (ATCC® 77103)

Applications: expression vectorvector permitting construction of fusion proteins  /  Depositors: JS Williams, JE Dixon

Designations pGEX-KG
Depositors JS Williams, JE Dixon
Biosafety Level 1
Distribution host: Escherichia coli HB101 (ATCC 33694)
Vector Information
Size (kb): 5.0000000000000000
Intact vector size: 5.000
Type of vector: plasmid
Cloning sites: BamHI SmaI EcoRI XbaI NcoI SalI XhoI SacI HindIII
Polylinker sites: EcoRI XbaI NcoI SalI XhoI SacI HindIII
Construction: pGEX-2T
Host range: Escherichia coli
Features (with orientation and position when available):
marker(s): ampR
promoter: tac
replicon: pMB1
repressor gene: lacIq
Vector: pGEX-KG (plasmid)
Promoters: Promoter tac
Construction: pGEX-2T
Construct size (kb): 5.0
Features: marker(s): ampR
promoter: tac
replicon: pMB1
repressor gene: lacIq
expression vector
vector permitting construction of fusion proteins
Restriction digests of the clone give the following sizes (kb): HindIII--5.0; BamHI--5.0; EcoRI--5.0; XbaI--5.0.
Expression vector permitting production of a fusion protein.
The glutathione S-transferase (GST) fusion protein can be purified by glutathione affinity chromatography, and the desired polypeptide released from the fusion product by thrombin.
Constructed from pGEX-2T by inserting an oligonucleotide at the EcoRI site which encodes the glycine "kinker" and additional restriction sites to facilitate cloning in all reading frames.
The order of the major features in this plasmid are: Ptac - GST - thrombin cleavage site - BamHI site - SmaI site - glycine "kinker" - multiple cloning site - ampR - pBR322 ori - lacIq.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C

Smith DB, Johnson KS. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67: 31-40, 1988. PubMed: 3047011

Guan KL, Dixon JE. Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal. Biochem. 192: 262-267, 1991. PubMed: 1852137

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