[Prokaryotic expression and purification of human histone-1 and its activity detection].
2011
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 Mr 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.
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