蛇毒蛋白原核表达包涵体复性研究进展

Recent Progress in Renaturation of Inclusion Bodies of Prokaryotically Expressed Snake Venom Proteins

外源基因在大肠杆菌中表达后常形成不溶性的无活性包涵体。包涵体的形成已经成为研究和应用活性蛋白质生产的主要障碍。然而,在合适的条件下,包涵体经过溶解、纯化、复性过程后可在体外重新折叠成有活性的蛋白质。迄今,已对蝰科、眼镜蛇科11种毒蛇的18个基因(包括金属蛋白酶、PLA2、β-银环蛇毒素、心脏素素、丝氨酸蛋白酶、神经生长因子、C-型凝集素等)成功进行了原核表达,采用稀释复性、透析复性和层析复性三种方法成功进行了包涵体复性。着重就蛇毒蛋白原核表达后包涵体复性所用的方法予以综述。

Insoluble,inactive inclusion bodies are often formed during prokaryotic expression of foreign genes.The formation of these deposits represents a major obstacle for the production of biologically active proteins for further investigation and application.However,after solubilization,isolation and refolding under suitable condition,inclusion bodies can convert to active proteins in vitro.So far,about eighteen snake venom proteins of eleven species in Viperidae and Elapidae（including metalloproteinases,phospholipase A2s,β-Bungarotoxin,cardiotoxins,serine proteases,nerve growth factors,C-type lectins） have been expressed in E.coli and successfully refolded by means of dilution,dialysis or chromatography.The refolding techniques for inclusion bodies of prokaryotically expressed snake venom proteins is summarized.