摘要
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.
出处
《生物工程学报》
CAS
CSCD
北大核心
2009年第3期428-434,共7页
Chinese Journal of Biotechnology
基金
国家科技支撑计划(No.2006BAD06A01)资助~~
关键词
PARP10
UV
组织表达谱
免疫沉淀
激光共聚焦
PARP10, ultraviolet ray, tissue expression profile, immuno-precipitation, laser confocal microscopy