pKM1000

pKM1000

Description: A Gateway™ destination vector for the production of recombinant proteins as fusions to the C-terminus of Thermatoga maritima maltodextrin-binding protein.

Origin: A derivative of pET11c (Novagen)

Replicon: ColE1

Antibotic Resistance: Ampicillin

Promotor: bacteriophage T7

Inducer: IPTG (1 mM)

Notes: The single cysteine in T. maritima MBP (residue 12) was changed to a serine by site-directed mutagenesis. Because it carries a gene (ccdB ) that encodes a lethal DNA gyrase poison, this vector must be propogated in an E. coli gyrA mutant such as DB3 or DB5 (Invitrogen), which is immune to the action of CcdB.

Literature Citations: Fox, J. D., Routzahn, K. M., Bucher, M. H., and Waugh, D. S. (2003). Maltodextrin-binding proteins from diverse bacteria and archaea are potent solubility enhancers. FEBS Lett. 537: 53-57. Wassenberg, D., Liefl, W., and Jaenicke, R. (2000). Maltose-binding protein from the hyperthermophilic bacterium Thermotoga maritima: Stability and binding properties. J. Mol. Biol. 295: 279-288.

Nucleotide Sequence:

Amino Acid Sequence: *

MKIEQTKLTI WSSEKQVDIL QKLGEEFKAK YGIPVEVQYV DFGSIKSKFL TAAPQGQGAD IIVGAHDWVG ELAVNGLIEP IPNFSDLKNF YDTALKAFSY GGKLYGVPYA MEAVALIYNK DYVDSVPKTM DELIEKAKQI DEEYGGEVRG FIYDVANFYF SAPFILGYGG YVFKETPQGL DVTDIGLANE GAVKGAKLIK RMIDEGVLTP GDNYGTMDSM FKEGLAAMII NGLWAIKSYK DAGINYGVAP IPELEPGVPA KPFVGVQGFM INAKSPNKVI AMEFLTNFIA RKETMYKIYL ADPRLPARKD VLELVKDNPD VVAFTQSASM GTPMPNVPEM APVWSAMGDA LSIIINGQAS VEDALKEAVD KIKAQIEKDQ TSLYKK AG...passenger protein.

* Amino acid residues encoded by the attR1 site in the Gateway™ cloning cassette are underlined.

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Description:	*A Gateway™ destination vector for the production of recombinant proteins as fusions to the C-terminus of Thermatoga maritima* maltodextrin-binding protein.**

Origin:	A derivative of pET11c (Novagen)

Replicon:	ColE1

Antibotic Resistance:	Ampicillin

Promotor:	bacteriophage T7

Inducer:	IPTG (1 mM)

Notes:	The single cysteine in T. maritima MBP (residue 12) was changed to a serine by site-directed mutagenesis. Because it carries a gene (ccdB ) that encodes a lethal DNA gyrase poison, this vector must be propogated in an E. coli gyrA mutant such as DB3 or DB5 (Invitrogen), which is immune to the action of CcdB.

Literature Citations:	Fox, J. D., Routzahn, K. M., Bucher, M. H., and Waugh, D. S. (2003). Maltodextrin-binding proteins from diverse bacteria and archaea are potent solubility enhancers. FEBS Lett. 537: 53-57. Wassenberg, D., Liefl, W., and Jaenicke, R. (2000). Maltose-binding protein from the hyperthermophilic bacterium Thermotoga maritima: Stability and binding properties. J. Mol. Biol. 295: 279-288.

Nucleotide Sequence: