Related Experiment Video
Updated: May 30, 2026
Purification of H3 and H4 Histone Proteins and the Quantification of Acetylated Histone Marks in Cells and Brain Tissue
Published on: November 30, 2018
[Prokaryotic expression and purification of human histone-1 and its activity detection]
Xiao-jie Xu1, Zhong-yi Fan, Ling-xue Wang
1Beijing Institute of Biotechnology, Beijing 100850, China.
Aim:
To construct the prokaryotic expression vector of human histone-1, obtain the purified GST-H1 protein, and detect its activity.
Methods:
Human histone-1 coding region was amplified from human mammary cDNA library, and was inserted into prokaryotic expression vector pGEX-KG. The recombinant plasmid pGEX-KG-H1 was transformed into E.coli Rossate. The expressed product was purified by GST-Sepharose 4B beads and identified by SDS-PAGE and Western blot analysis.
Results:
The DNA fragment of about 650 bp was successfully amplified by PCR, cloned into pGEX-KG, and identified by sequencing. The recombinant protein of about M(r); 52 000 was successfully induced, purified and tested well by Kinase assay.
Conclusion:
The recombinant protein of GST-H1 is obtained successfully, which lay the foundation for further research on cell cycle control.
