蛋白 G 的优化 C2 结构域的特性：寻找其额外的鸡 IgY 结合能力。

Characterization of the optimized C2 domain of protein G: finding its additional chicken IgY-binding ability.

机构信息

College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, People's Republic of China.

出版信息

Biotechnol Lett. 2013 Sep;35(9):1441-7. doi: 10.1007/s10529-013-1221-7. Epub 2013 May 21.

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.

摘要

链球菌蛋白 G 的 C2 结构域是一个小的（55 个残基）肽，具有免疫球蛋白结合活性。经过密码子优化后，该基因被分成四个寡核苷酸片段，并通过重叠 PCR 进行扩增。重组质粒 pET30a-C2 转化入大肠杆菌 Rosetta（DE3）PLysS 进行表达。经 Ni-NTA 纯化后，通过 Western-blotting、Dot-ELISA 和 ELISA 鉴定融合蛋白。His 标记的 C2 与人和兔、牛、猪、山羊、鼠或豚鼠 IgG 结合，但与鹅、鸭、野鸭、火鸡和丹顶鹤 IgY 没有亲和力。然而，它与鸡 IgY 的亲和力与豚鼠 IgG 相当。因此，C2 结构域可能为从各种哺乳动物中纯化和检测免疫球蛋白 G 提供了理想的材料。