关键词: PIASxβ, pCMV-Myc载体, 重组质粒, 基因表达

Abstract:

Abstract:Objective To construct the recombinant plasmid pCMV-Myc-PIASXβ and express the fusion protein in mammalian cells.Methods  PIASxβ fragment was digested from the original vector pGADT7 with SalⅠand NotⅠ,and then was inserted into the targeted pCMV-Myc vector by the recombinant DNA technique.After identification,the recombinant plasmid was transfected　into CHO cells.The expression of recombinant Myc-PIASxβ fusion protein was detected by Western blotting.
Results By the restriction enzyme digestion,fragment purification,ligation and transformation,the recombinant plasmid was obtained.The right recombinant plasmid pCMV-Myc-PIAS3 was identified with enzyme digestion and sequencing.By EcoRⅠ digestion analysis,pCMV-Myc-PIASxβ showed a 5641 bp band.By XbaⅠdigestion analysis,pCMV-Myc-PIASxβ showed two expected band of 3291 bp and 2349 bp.A specific protein expression band at 68 000 (PIASxβ fusion protein) was showed in Western blotting,which matched recombinant plasmid.
Conclusion The recombinant plasmid of pCMV-Myc-PIASxβ is sucssesefully constructed,which provids a good tool for further function study on PIAS family．

Key words: PIASxβ, pCMV-Myc vector, recombinant plasmid, genen expression