HE Yujuan, et al. The expression of open reading frame of unknown KH gene from K562 cells in E.coli[J]. Journal of Chongqing Medical University, 2004,(2):124-126.
HE Yujuan, et al. The expression of open reading frame of unknown KH gene from K562 cells in E.coli[J]. Journal of Chongqing Medical University, 2004,(2):124-126. DOI: 10.13406/j.cnki.cyxb.2004.02.002.
Objective:To construct the prokaryotic expression system of open reading frame of KH gene (KH-ORF)
an unknown gene in K562 cells.Methods:The 225bp KH-ORF cDNA was cleaved from plasmid pGEM-T-easy-KH-ORF with EcoRI and SalI and subcloned into the pCI-neo Mammalian Expression Vector cutted with the same restriction endonucleases.The recombinant expression plasmid was transfected into BL21 (DE3) plysS by hot-shock assay.Induced by IPTG
the recombinant protein product was tested by SDS-PAGE.Results:24kD KH-ORF product was obtained as inclusion bodies in BL21 (DE3) plysS after the expression vector had been induced by IPTG for 30 minutes Conclusion :The construction of prokaryotic expression system of KH- ORF is successful.