新生与成年土拨鼠肝组织中部分差异表达的基因比较分析
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艾滋病和病毒性肝炎等重大传染病防治


To compare and analyse some parts of differently expressed genes in liver between neonatal and adult woodchucks
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    摘要:

    目的 新生土拨鼠感染土拨鼠肝炎病毒后,大部分发展为慢性肝炎,而成年土拨鼠感染后则多发生急性自限性肝炎[1]。本实验目的就是寻找其肝组织中可能导致这种预后差异的关键基因。 方法 采用全基因组表达谱芯片技术,对比新生与成年小鼠肝组织基因表达差异,选取目的基因,再通过多个物种序列比对,设计简并引物,在土拨鼠肝组织cDNA中扩增对应基因,测序,再次设计引物,进行实时荧光定量PCR。 结果 与新生土拨鼠相比,成年土拨鼠肝细胞中与钙离子重吸收相关基因DNM1(Dynamin 1)、DNM3(Dynamin 3)及Prkcc(Protein kinase C,gamma)表达率明显升高,分别上升2.65?.25倍,1.90?.34倍,2.94?.54倍。 结论 在钙离子重吸收通路中,以上三个在两组动物肝脏中表达差异最明显的基因,在新生组与成年组土拨鼠之间也有明显差异。此类基因造成肝细胞内钙离子浓度的差别,间接影响其中肝炎病毒的复制。这种表达差异很可能是导致两个年龄段动物感染土拨鼠肝炎病毒后转归不同的原因之一。

    Abstract:

    Objective To search for some key genes which account for the different sequelae(chronic woodchuck hepatitis and acute self-limiting woodchuck hepatitis) of woodchuck hepatitis virus infection between neonatal and adult woodchucks. Methods Applying the genome-wide expression microarray, some related genes were selected. With the degenerate primers designed on the iCODEHOP from clustal of some different species, some products were amplified from the woodchuck cDNAs. After the products sequencing done, some accurately designed primers generated from the Primer Premier 5.0, Real time quantitative PCR performed ultimately. Results Compare to the neonatal woodchuck, calcium reabsorption related gene, in the adult woodchuck liver cells DNM1 ( Dynamin 1 ), DNM3 ( Dynamin 3 ) and Prkcc ( Protein kinase C, gamma ) expression rate increased significantly, by 2.65?0.25 times, 1.90 ?.34 times, 2.94 ?.54 times. Conclusion In the endocrine and other factor-regulated calcium reabsorption pathway, these three genes have the most significant changes in mice, as well as in woodchucks at the two time points. Those genes make contribute to the differential calcium concentration, and promote woodchuck hepatitis virus replication indirectly. In a word, these genes could have an effect on the sequelae of woodchuck hepatitis virus infection at different ages.

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王响.新生与成年土拨鼠肝组织中部分差异表达的基因比较分析[J].中国比较医学杂志,2013,23(2).

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  • 收稿日期:2012-12-24
  • 最后修改日期:2012-12-29
  • 录用日期:2013-01-09
  • 在线发布日期: 2013-03-05
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