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作 者:于孟斌[1,2] 赵莉霞[1] 杨予涛[1] 杨志新[1] 张莹莹[1] 朱恒奇[1] 周晓巍[1] 黄培堂[1]
机构地区:[1]军事医学科学院生物工程研究所,北京100071 [2]防化指挥工程学院生物防护教研室,北京102205
出 处:《生物工程学报》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~~
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.
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