Recombination and expression of soluble human CD59 in E. Coli
Abstract：Objective To obtain soluble recombinant human CD59 with high activity. Methods The gene sequence which codes natural CD59 extracellular portion containing 77 amino acids was amplified with PCR. The fragment was cloned into prokaryotic expression vector PinPoint Xa-3. The recombinant protein expression was induced by IPTG and biotinylated products were purified by avidin resin. The activity of purified recombinant CD59 was evaluated by reactive assay in vitro. Resuits In PinPoint Xa-3 expression system, we obtained recombinant CD59 with a molecular wright of 22kDa. The thin layer gel scanning analysis showed that about 7% of total cell protein is newly expressed fusion protein. SDS-PAGE assay showed that the product affinity-purified by avidin resin has high purity. The reactive assay revealed that the purified protein had the activity of inhibiting hemolysis. Conclusion The fusion protein has the activity of natural CD59 to some extent.
Keywords：recombinant CD59fusion proteinexpression
Publication Date：2000-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 96-98 )