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

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.