Prokaryotic expression of human Hlx gene from the cord blood mononuclear cells and immunogenicity analyzing of its fusion protein
DAI Xiaoli
WANG Shengjun
SU Zhaoliang
PENG Sufang
LI Yazhen
XUE Yuan
HE Zhiqiang
CHEN Jianguo
SHAO Qixiang
HUANG Xinxiang
XU Huaxi
Abstract：Objective To clone and express human Hlx in M15 strain of E.coli using pQE30 vector for preparing polyclonal antibody against Hlx.Methods The open reading frame of full length Hlx gene was cloned from the mononuclear cells of human cord blood by RT-PCR.Recombinant expression plasmid pQE30/Hlx,obtained by transfecting Hlx into pQE30 vector,was expressed in E.coli M15.Western blotting was used to detect the expression of Hlx soluble fusion protein with 6×His tag.The protein was purified by Ni~(2+)-MAC column,and then used to immunize rabbit for preparing polyelonal antibody against Hlx.Results The full length of Hlx gene open reading frame containing 1 467 bp and encoding 488 amino acids was cloned successfully.The human Hlx protein was expressed and purified.Polyclonal antibody against human Hlx was gained,which possesses high reactivity and specificity approved by ELISA and Western blotting,respectively.Conclusion Our group constructed prokaryotic expression plasmid pQE30/Hlx successfully,gained high purified protein,as well as prepared high potency and specific polyclonal antibody against human Hlx,which make an initial foundation for further research on the function of Hlx.
Keywords：Hlxprokaryotic expressionprotein purifypolyclonal antibody
Publication Date：2009-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 625-628 )