Cloning and expression of soluble recombinant protein comprising the extracellular domain of the human type I interferon receptor 2c subunit (IFNAR-2c) in E. coli |
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| Authors: | Sun-Ok Yoon Rosario D.C. Hirata Ana C.R. da Silva Nga Y. Nguyen Mario H. Hirata |
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| Affiliation: | (1) Department of Clinical and Toxicological Analysis, Faculty of Pharmaceutical Science, Institute of Chemistry, University of São Paulo, São Paulo, SP, 05508-900, Brazil;(2) Facility for Biotechnology Resources, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD, USA;(3) Department of Clinical and Toxicological Analysis, Faculty of Pharmaceutical Science, Institute of Chemistry, University of São Paulo, São Paulo, SP, 05508-900, Brazil |
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| Abstract: | An optimized procedure was developed for production of the extracellular domain encoding amino acids 1–243 of the human type I interferon receptor 2c subunit (IFNAR-2c) as a fusion protein with glutathione S-transferase (GST-IFNAR2cEC) in E. coli to obtain active, soluble protein. Induction of protein expression at 37 °C resulted in a formation of inclusion body. Co-expression with bacterial chaperones, GroEL and GroES, failed to support the folding of GST-IFNAR2cEc under IPTG induction at 37 °C. Expression induced at 25 °C decreased the inclusion body formation up to 62% and cell disruption by a French press provided higher recovery of the recombinant protein than cell disruption by sonication. |
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| Keywords: | chaperones recombinant soluble protein type I interferon receptor 2c subunit |
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