• Yuan-ming Wu, Xiao-nan Zhang, Xiao-hong Xin, Jun-jie Zhang, Nan-chun Chen, and Su-min Chen.
• Center for DNA Typing, Fourth Military Medical University, Xi'an, China. wuym@fmmu.edu.cn
• Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Jun 1; 24 (6): 560-3.

AimTo prepare a soluble human Era(hEra) protein and to measure its bioactivity.MethodsHuman era cDNA gene from pUC19 plasmid was subcloned into the expression plasmid pMAL-p2x. pMAL-hEra was transducted to E.coli TB1 and the strain was induced by isopropyl beta-D-thiogalactopyranoside (IPTG).ResultsThe expressed MBP-fused protein existed in a soluble form. The fused protein made up 23.9% of the total cell lysate. It was purified by amylose affinity chromotography and digested with Factor X. Although the fused segment was dissected, the remained hEra protein was unstable in the solution with the passage of time. The activity assay showed that hEra was a GTPase that could bind GTP and hydrolyze GTP to GDP.ConclusionHuman Era protein can be expressed in a soluble form and it has been proved to be a kind of G protein by the experiments in vitro. The study is important to further research into the function of human era gene.

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