关键词: RUNX3, 载体, 蛋白诱导, 蛋白表达

Abstract:

Abstract:Objective
To construct the prokaryotic expression vectors of overall length and different domains of RUNX3 and to identify the expressions of their recombinant proteins.Methods The coding sequence of overall length and different domain truncations of RUNX3 were amplified from the pcDNA3.1-myc-RUNX3 plasmid  by PCR method and inserted into glutathione transferase(GST) fusion expression vector  pGEX-4T-2 through EcoRⅠand BamHⅠ restriction enzyme sites.Then they were transformed into E.coli BL21 and the fusion proteins were induced  and identified by SDS-PAGE electrophoresis.Results The vector fragments FL (1 245 bp),ΔRunt(843 bp),Nter(159 bp),Runt(402 bp), and Cter (684 bp) which were  consistent with the expected fragments were obtained after double enzyme digestion of EcoRⅠand BamHⅠ.The SDS-PAGE electrophoresis result showed that the relative molecular mass of GST-RUNX3 truncated fusion proteins of overall length and different domains of  RUNX3 (GST-FL,GST-Runt,GST-Nter and GST-Cter)were 72 000,40 000,32 000, and 51 000,respectively.Conclusion GST-tagged prokaryotic expression vectors overall length and   different truncated regions of RUNX3  are constructed  and their recombinant proteins are expressed successfully.

Key words: RUNX3, vector, inducing of protein, expression of protein