关键词: <, p>, 人, NEDD8, 原核表达, 多抗<, /p>

Abstract: Abstract: In the study，human nedd8 gene ( HN8) was synthesized and subcloned into pMD19-T simple vector．After sequencing，HN8 gene was transferred into the prokaryotic expression vector，pCold-TF． The final construct is designated as pCold-HN8． The plasmid was transformed into E． coli BL21 ( DE3) competent cells． Induced with Isopropyl β-D-1-thiogalactopyranoside ( IPTG) ，the bacteria were grown in LB broth at 16℃ for 24 hours． The products were sonicated and then purified by His-bind purification kit． The purified protein was used to immunize three rabbits to prepare polyclonal antibody． The results showed that the fused protein were soluble expressed with a size of 64 kDa by SDS-PAGE analysis． Both indirect immunofluorescence and Western blotting assay，it confirmed the polyclonal antibody could recognize the HN8 protein． This antibody provides a useful tool for a further study of the neddylation modification by human NEDD8．

Key words: <p>human, NEDD8, prokaryotic expression, polyclonal antibody</p>