Construction Of The Murine Interleukin-10 Plasmid Enhanced By Adeno-associated Virus And Its Expression In Murine Cardiomyocytes

[Objective] 1 Construct the murine interleukin-10 plasmid enhanced by ITR of adeno-associatedvirus and improve ITR could increase the expression of mIL-10 in murinecardiomyocytes. [methods]1 To insert inverted terminal repeats(ITR) of adeno - associated virus(AAV) intopORF5-mIL-10 plasmid and to construct the murine interleutin-10 plasmid enhanced by ITR.2 pORF5-mIL-10 and pORF-ITR-mIL-10 were transfected into cardiomyocytes separately induced by lipofectin3 mRNA of mIL-10 extracted by Trizol one step method was detected with RT-PCR.4 Exame the expression of mIL-10 in the culture supermanant by ELIS A. [Results]1 Confirm to construct the pORF-ITR-mIL-10 plasmid successfully by double -digestion2 24h, 48h, 72h, 96h. 120h after transfection , the target bars were seen in RT-PCR;3 ELIS A showed the expressive quantity of pORF-ITR-mIL-10 was higher than that of pORF5-mIL-10.[Conclusion] pORF-ITR-mIL-10 can increase the expression level of mIL-10 in murinecardiomyocytes.