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HeLa/GFP—H2B细胞系的构建及对其有丝分裂进程的动态观察 被引量:3

Construction and Mitosis Observation of HeLa/GFP—H2B Cell
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摘要 准确的染色体分离依赖于有丝分裂过程的精确调控,包括有丝分裂的时间,及纺锤体检查点的正确调控等。通过动态观察有丝分裂染色体的运动可对上述研究进行精确定量。结果显示,利用逆转录病毒系统成功构建了稳定融合表达绿色荧光蛋白GFP—H2B的HeLa细胞系,结合细胞同步化方法,建立了一套利用活细胞荧光共聚焦显微镜观察HeLa细胞有丝分裂的实验体系。 Accurate chromosome segregation depends on the precise regulation of mitotic progression,including the timing of mitosis and proper regulation of spindle assembly checkpoint are getten.Though live-cell observation movement of chromosome,we could accurately quantify the procedure above.Here,though retroviral infection system we get stable expressed GFP-H2B HeLa cell line.Together with synchronizing cells to mitosis,a technique is established that observing mitosis though time-lapse confocal scanning laser microscopy.
出处 《科学技术与工程》 2011年第21期5150-5153,5157,共5页 Science Technology and Engineering
基金 国家自然科学基金(30871234)资助
关键词 有丝分裂 细胞同步化 活细胞观察 mitosis cell synchronization live-cell imageing
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  • 1Meraldi P,Draviam V M, Sorger P K. Timing and checkpoints in the remalation of mitotic oroaression. Dev Ce11.2004:7 ( 1 ) : 45-60.
  • 2Neumann B,Walter T, Heriche J K,et al. Phenotypic profiling of the human genome by time-lapse microscopy reveals cell division genes. Nature,2010 ;464 (7289) : 721-727.
  • 3Sonnichsen B, Koski L B, Walsh A ,et al. Full-gertonae RNAi profiling of early embryogenesis in Caenorhabditis elegans. Nature,2005; 434 (7032) : 462-469.
  • 4Neumann B, Held M, Liebel U,et al. High-throughput RNAi screening by time-lapse imaging of live human cells. Nat Methods ,2006 ;3 (5) : 385-390.
  • 5Walczak C E, Cai S, Khodjakov A. Mechanisms of chromosome behaviour during mitosis. Nat Rev Mol Cell Biol, 2010; 11 (2) : 91-102.
  • 6Li Y, Gorbea C, Mahaffey D. MAD2 associates with the cyclosome/ anaphase-promoting complex and inhibits its activity. Proceedings of the National Academy of Sciences of the United States of America, 1997;94 (23) : 12431-12436.
  • 7Reddy S K, Rape M, Margansky W A, et al. Ubchl0-Ubiquitination by the anaphase-promoting complex drives spindle checkpoint inactivation. Nature,2007;466 (7138) : 921-925.
  • 8Garnett M J, Mansfeld J, Godwin C. UBE2S elongates ubiquitin chains on APC/C substrates to promote mitotic exit. Nat Cell Bio1,2009 : 11(11) : 1363-1369.
  • 9Lippincott-Schwartz J, Patterson G H. Development and use of fluorescent protein markers in living cells. Science, 2003 :300 ( 5616 ) :87-91.

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  • 1Zhang P J, Zhan J, Zhang X M, et al. CUE domain containing 2 regulates degradation of progesterone receptor by ubiquitin Cproteasome. The EMBO journal, 2007 ; 26 (7) : 1831-1842.
  • 2Pan X, Zhou T, Tai Y H, et al. Elevated expression of CUEDC2 protein confers endocrine resistance in breast cancer. Nature Medicine, 2011; 17:708-714.
  • 3Gao Y F, Li T, Chang Y, et al. Cdkl-phosphorylated CUEDC2 promotes spindle checkpoint inactivatima and chromosomal instability. Nat Cell Biol, 2011 ; 13(8) :924-933.
  • 4Luo Xiuquan, Huang Ying, Sheikh M S, et al. Cloning and charac- terization of a novel gene PDRG that is differentially regulated by p53 and ultraviolet radiation. Oncogene, 2003, 22:7247-7257.
  • 5Jiang Lingyan, Luo Xiuquan, Shi Jingxue, et al. PDRG1, a novel tumor marker for muhiple malignancies that is selectively regulated by genotoxic stress. Cancer Biology & Therapy , 2011, 11 ( 6 ) : 567-573.
  • 6Li H Y, Liu H, Zhang X M, et al. Deactivation of the kinase IKK by CUEDC2 through recruitment of the phosphatase PP1. Nature Immu- nology ,2008 , 9 (5) : 533-541.
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