title: Characterization of the optimized C2 domain of protein G: finding its additional chicken IgY-binding ability.
abstract: The C2 domain of streptococcal protein G is a small (55 residue) peptide with immunoglobulin-binding activity. Following codon optimization, the gene was divided into four oligonucleotide fragments and amplified by overlap PCR. The recombinant plasmid pET30a-C2 was transformed into Escherichia coli Rosetta (DE3) PLysS for expression. After purification by Ni-NTA, the fusion protein was identified by western-blotting, Dot-ELISA and ELISA. His-tagged C2 bound to human, rabbit, cattle, pig, goat, mouse or guinea pig IgG had no affinity for goose, duck, wild duck, wild turkey and red-crowned crane IgY. Its affinity for chicken IgY, however, was comparable to that of guinea pig IgG. The C2 domain may therefore provide an ideal material for the purification and detection of immunoglobulin G from various mammals.
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, People's Republic of China. caoys87@sina.com.cn
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Cao
authors.forename: Di
authors.initials: D
authors.lastname: Li
authors.forename: Chong
authors.initials: C
authors.lastname: Cao
authors.forename: Mingchun
authors.initials: M
authors.lastname: Gao
authors.forename: Runxiang
authors.initials: R
authors.lastname: Zhang
authors.forename: Bo
authors.initials: B
authors.lastname: Ma
authors.forename: Junwei
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002645:Chickens; D004926:Escherichia coli / Q000235:genetics; D007136:Immunoglobulins / Q000378:metabolism*; D010957:Plasmids; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; C016512:IgY; D007136:Immunoglobulins; D011994:Recombinant Proteins