Expression and single-step purification of mercury transporter (merT) from Cupriavidus metallidurans in E. coli.
The mercury transporter, merT, from Cupriavidus metallidurans was cloned into pRSET-C and expressed in various E. coli hosts. Expression of merT gene failed in common expression hosts like E. coli BL21(DE3), E. coli BL21(DE3)pLysS and E. coli GJ1158 due to expression induced toxicity. The protein was successfully expressed in E. coli C43(DE3) as inclusion bodies. The inclusion bodies were solubilized with Triton X-100 detergent. The detergent solubilized protein with N-terminal His-tag was purified in a single-step by immobilized metal affinity chromatography with a yield of 8 mg l(-1).
Senthil
K
K
Centre for Biotechnology, Anna University, Chennai 600025, India.
Gautam
P
P
D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification*; D027682:Cation Transport Proteins / Q000235:genetics* / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D049921:Cupriavidus / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology; D015870:Gene Expression; D002479:Inclusion Bodies / Q000201:enzymology; D050296:Microbial Viability; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000633:toxicity
D001426:Bacterial Proteins; D027682:Cation Transport Proteins; D011993:Recombinant Fusion Proteins; C094115:mercury transport protein, bacterial