人可溶性CD59的原核表达及其生物学活性的初步研究

Expression of Human Soluble CD59 Gene in Ecoli and Preliminary Identification of Its Activity

重组人可溶性CD5 9(rhsCD5 9)。将rhsCD5 9的基因克隆于表达载体PinPointXa 3的多克隆位点 ,构建成生物素化sCD5 9融合蛋白表达载体pPinPointXa sCD5 9,将此表达载体转化大肠杆菌JM10 9,IPTG诱导表达后 ,收集裂解菌体 ,取上清过已处理好的SoftLink亲和素树脂柱 ,分段收集融合蛋白 ,以Xa切割后再过SoftLink柱获得rhsCD5 9,初步鉴定rhsCD5 9的生物活性。rhsCD5

Human soluble CD59(hsCD59) gene coding the soluble part of Nterminus with 70 amino acids residues was recombined The gene was cloned into the HindⅢ/BglⅡ site of PinPoint Xa3 and placed under the transcriptional control of the tac promoter followed by the codons of 126 amino acids biotinbinding protein The biotinylated sCD59,a fusion protein,was expressed in Ecoli,JM 109 ,at 37℃ with 500mmol IPTG induction for 4h The biotinbinding protein was purified with SoftLink Avidin Resin,digested with factor Xa and purified once more with SoftLink Avidin Resin and the rhsCD59 was collected The effect of the sCD59 on sublytic damage of the complement MAC on the glomerular endothelia cells was observed