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Expression and single-step purification of mercury transporter (merT) from Cupriavidus metallidurans in E. coli.
- Source :
-
Biotechnology letters [Biotechnol Lett] 2010 Nov; Vol. 32 (11), pp. 1663-6. Date of Electronic Publication: 2010 Jun 24. - Publication Year :
- 2010
-
Abstract
- 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).
- Subjects :
- Chromatography, Affinity methods
Cupriavidus genetics
Escherichia coli genetics
Escherichia coli physiology
Gene Expression
Inclusion Bodies enzymology
Microbial Viability
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins isolation & purification
Recombinant Fusion Proteins toxicity
Bacterial Proteins genetics
Bacterial Proteins isolation & purification
Cation Transport Proteins genetics
Cation Transport Proteins isolation & purification
Cupriavidus enzymology
Subjects
Details
- Language :
- English
- ISSN :
- 1573-6776
- Volume :
- 32
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- Biotechnology letters
- Publication Type :
- Academic Journal
- Accession number :
- 20574829
- Full Text :
- https://doi.org/10.1007/s10529-010-0337-2