PARP10组织分布、相互作用蛋白及UV应激反应的分析

Analysis of PARP10 tissue expression profile, interactive protein and UV stress reaction

根据GenBank中公布的人多聚腺苷酸二磷酸核糖聚合酶10(PARP10)cDNA序列,设计并合成一对特异性引物,通过RT-PCR扩增293FT细胞mRNA,获得PARP10cDNA。将获得的cDNA克隆到pCMV-Myc和pEGFP-C1中,用免疫沉淀和激光共聚焦实验验证了PARP10和β-actin存在相互作用。然后,通过RT-PCR法发现该基因在小鼠体内表达的组织分布,组织表达谱显示该蛋白在各组织中均有表达;Westernblotting分析表明,UV造成的细胞损伤能够引起PARP10表达水平的增高。PARP10组织表达谱的确定,与β-actin相互作用的验证以及对UV的应激反应都为进一步研究PARP10的生物学功能奠定了基础。

One pair of primers were designed and synthesized based on the cDNA sequence encoding Homo sapiens poly （ADP-ribose） polymerase family, member 10 （PARP10） reported on the GenBank. The cDNA sequence encoding PARP10 was cloned from 293FT cell by RT-PCR. Then the RT-PCR product was cloned into pCMV-Myc and pEGFP-C1 plasmids. The interaction between PARP10 and β-actin was identified through immuno-precipitation and laser confocal microscopy. Extensive expression of PARP10 in mouse tissues was confirmed by RT-PCR. Besides, Western blotting analysis indicated that cell injury caused by UV treatment could promote the expression of PARP 10. The results in this paper would benefit further study of PARP 10.