title: Production of Recombinant CCN2 Protein in Escherichia coli.
abstract: Recombinant proteins are important tools for understanding molecular functions in vitro. Recent progress in the generation of recombinant proteins is amazing. However, when we plan to produce them, we should choose the best method according to the nature and the use of the target recombinant protein. Degradation and mis-folding are major problems in producing active recombinant CCN2. The method shown in this chapter describes the appropriate conditions under which we can produce CCN2 and its truncated fragments in Escherichia coli.
authors.affiliation: Advanced Research Center for Oraland Craniofacial Sciences, Okayama University Dental School/Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1, Shikata-cho, Kita-ku, Okayama, 700-8525, Japan.
authors.forename: Eriko
authors.initials: E
authors.lastname: Aoyama
authors.affiliation: Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Hattori
authors.affiliation: Advanced Research Center for Oral and Craniofacial Sciences, Okayama University Dental School/Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan.|Department of MembraneBiochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kubota
authors.affiliation: Advanced Research Center for Oraland Craniofacial Sciences, Okayama University Dental School/Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1, Shikata-cho, Kita-ku, Okayama, 700-8525, Japan. takigawa@md.okayama-u.ac.jp.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Takigawa
mesh_terms: D055511:CCN Intercellular Signaling Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D011993:Recombinant Fusion Proteins
chemical_list: D055511:CCN Intercellular Signaling Proteins; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins
keywords: Escherichia coli (E. coli); FLAG peptide; FLAG-tag; Histidine6-tag; Imidazole; Ni-NTA agarose gel