Related Experiment Video
Updated: Jul 4, 2026
Recombinant Protein Expression for Structural Biology in HEK 293F Suspension Cells: A Novel and Accessible Approach
Published on: October 16, 2014
[Soluble expression, purification and bioactivity of recombinant human Era protein]
Yuan-ming Wu1, Xiao-nan Zhang, Xiao-hong Xin
1Center for DNA Typing, Fourth Military Medical University, Xi'an, China. wuym@fmmu.edu.cn
Aim:
To prepare a soluble human Era(hEra) protein and to measure its bioactivity.
Methods:
Human 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).
Results:
The 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.
Conclusion:
Human 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.
