Küster E, Luesink E J, de Vos W M, Hillen W
Lehrstuhl für Mikrobiologie, Friedrich-Alexander-Universität Erlagen-Nürnberg, Germany.
FEMS Microbiol Lett. 1996 Jun 1;139(2-3):109-15. doi: 10.1111/j.1574-6968.1996.tb08188.x.
The catabolite control protein CcpA from Bacillus megaterium was overproduced as a fusion protein to a 6xhis affinity tag and purified to homogeneity. Polyclonal antibodies of high affinity and specificity were raised against the purified protein. The serum did not crossreact with purified Lac repressor despite the fact that CcpA and LacI belong to the same protein family. Using this antiserum we identified proteins that share antigenic determinants with CcpA in many Gram-positive bacteria, including bacilli, staphylococci, lactic acid bacteria, and some actinomycetes.
巨大芽孢杆菌的分解代谢物控制蛋白CcpA作为与6xhis亲和标签的融合蛋白过量表达,并纯化至同质。针对纯化后的蛋白制备了具有高亲和力和特异性的多克隆抗体。尽管CcpA和LacI属于同一蛋白家族,但该血清与纯化的乳糖阻遏蛋白没有交叉反应。利用这种抗血清,我们在许多革兰氏阳性细菌中鉴定出了与CcpA具有共同抗原决定簇的蛋白,这些细菌包括芽孢杆菌、葡萄球菌、乳酸菌和一些放线菌。