High efficient expression of GST-hureb1 and generation of anti hureb1 antibody / 中国免疫学杂志

ABSTRACT

Objective:To construct high level prokaryotic cell expression vector carrying human upstream regulatory element binding protein1 (hureb1) gene for producing GST hureb1 fusion protein and generating anti hureb1 antibody Methods: After the Xho I/Not I fragment from hureb1 open reading frame was recombined into the prokaryotic expression vector pGEX 4T 2, the vector was transformed into E coli BL21(DE3) strain and induced with IPTG to express the fusion protein, GST hureb1 The crudely isolated fusion protein was applied to immunize rabbit and mouse for generating anti hureb1 polyclonal antibody and monoclonal antibody respectively Results:The prokaryotic expression vector expressing GST hureb1 fusion protein was constructed and induced to produce high level GST hureb1 that was detected up to 33 45% of the total bacterial protein expressed The GST hureb1 fusion protein immunizing animals generated high titer and specific anti hureb1 antibodies Conclusion:Recombinant hureb1 protein and anti hureb1 antibodies can be used to analyze the biological functions of hureb1