Expression, purification and application of staphylococcal protein A fused to SUMO
10.3760/cma.j.issn.0254-5101.2010.07.002
- VernacularTitle:金黄葡萄球菌蛋白A与SUMO标签的融合表达及应用研究
- Author:
Xuehui GAO
;
Jiechao YIN
;
Guopeng SUN
;
Wenfei WANG
;
Deshan LI
- Publication Type:Journal Article
- Keywords:
Staphylococcal protein A;
SUMO;
Fusion protein;
Antibody purification
- From:
Chinese Journal of Microbiology and Immunology
2010;30(7):597-602
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone the full length staphylococcal protein A(SPA) gene from Staphylococcus aureus (ATCC6538), and subsequently study the gene structure and antibody binding ability.Methods The full length and the functional region of the SPA gene were cloned into pHisSUMO vector respectively, and expressed in E. coli. The full length and the functional fragment of the SPA protein were detected for antibody binding ability and stability. The functional fragment of the SPA protein fused with SUMO was coupled to the CNBr-activated agarose for antibody purification from rabbit serum. Results A variant of the full length SPA gene was cloned, which has been submitted to GenBank (the accession number is EU695225). Two fusion proteins had the same antibody binding ability as the untagged SPA protein. However, the formers was more stable than the latter at the tested conditions. SUMO-SPA conjugated-agarose kept high efficiency for antibody binding. Conclusion To our knowledge, the full length SPA gene of S.aureus(ATCC6538) is a novel variant. The SUMO tag can improve the stability of the functional region of the SPA protein without damaging the antibody binding ability. This fusion protein has been used for antibody purification successfully.
