SpiC/SsaM蛋白质复合物的原核表达及纯化

沙门氏菌毒力岛2中spiC所编码的效应蛋白在沙门氏菌得以于宿主吞噬性细胞内的存活中起到重要作用．SpiC与位于相同操纵元的SsaM特异结合，暗示了SpiC／SsaM蛋白质复合物参与转运元的形成并调控其他效应蛋白的分泌．为了从蛋白质结构角度研究SpiC／SsaM复合物的生物学功能，原核体系中共表达了SpiC和SsaM．与SpiC形成稳定的复合物，改善了SsaM单独表达不可溶的情况．胰蛋白酶限制性酶切实验显示：相对单独存在于溶液中的SpiC，形成复合物的SpiC更加稳定．得到大量高纯度的胰蛋白酶处理后的SpiC／SsaM复合物，并进行了结晶试验，为后续结构学研究奠定了基础．

Prokaryotic expression and purification of SpiC/SsaM protein complex

The gene of a novel protein known as SpiC located on Salmonella Pathogenicity Island 2 is required for the survival of Salmonella within macrophages.The fact that another gene ssaM is also located within the same operon of spiC suggests that a complex involving SsaM and SpiC may function in the formation of translocators and control other effector proteins' order of secretion.To facilitate the structural study of this complex,SpiC and SsaM were cloned and expressed in the prokaryotic expression system.Altho...