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小鼠胎儿胸腺细胞系中基因表达的序列分析

Serial analysis of gene expression in murine fetal thymocyte cell lines.

作者信息

Zhao Feng-Qi, Sheng Zong-Mei, Tsai Mark M, Hubbs Alan E, Wang Ruixue, O'Leary Timothy J, Izon David J, Taubenberger Jeffery K

机构信息

Molecular Pathology Division, Department of Cellular Pathology and Genetics, Armed Forces Institute of Pathology, Rockville, MD 20850, USA.

出版信息

Int Immunol. 2002 Dec;14(12):1383-95. doi: 10.1093/intimm/dxf103.

Abstract

FTL-1, -3 and -10 are three murine day 14 fetal thymocyte cell lines produced in order to model developmental stages within early (CD3-CD4-CD8-) thymocyte differentiation. In this study, we used the serial analysis of gene expression (SAGE) method to perform a systematic analysis of transcripts present in these three cell lines. A total of 77,313 SAGE tags were sequence identified from the three cell lines, representing 24,645 unique transcripts. Differentially expressed mRNA transcripts representing different gene classes were identified, including T cell functional genes, cytokine receptors, adhesion molecules and transcription factors. These results may serve as a model of the transcriptome of early thymocyte differentiation. A large number of unknown expressed sequence tags were also found to be differentially expressed. In order to validate the SAGE data, selected differentially expressed transcripts identified by SAGE were analyzed by quantitative RT-PCR in normal murine double-negative stage DN1-4 thymocytes. Expression of the transcription factors RUNX2 and PHD finger protein 2 and of the IGF type 1 receptor was shown to have differentially regulated expression patterns in sorted DN1-4 cells. These genes, and others identified by this analysis, are likely to play important roles in the development of T cells.

摘要

FTL-1、-3和-10是三种小鼠第14天胎儿胸腺细胞系,构建这些细胞系是为了模拟早期(CD3-CD4-CD8-)胸腺细胞分化过程中的发育阶段。在本研究中,我们使用基因表达序列分析(SAGE)方法对这三种细胞系中存在的转录本进行了系统分析。从这三种细胞系中总共测序鉴定出77313个SAGE标签,代表24645个独特的转录本。鉴定出了代表不同基因类别的差异表达mRNA转录本,包括T细胞功能基因、细胞因子受体、黏附分子和转录因子。这些结果可作为早期胸腺细胞分化转录组的模型。还发现大量未知的表达序列标签存在差异表达。为了验证SAGE数据,通过定量RT-PCR在正常小鼠双阴性阶段DN1-4胸腺细胞中分析了SAGE鉴定出的选定差异表达转录本。结果显示,转录因子RUNX2和PHD指蛋白2以及胰岛素样生长因子1型受体在分选的DN1-4细胞中具有差异调节的表达模式。这些基因以及通过该分析鉴定出的其他基因可能在T细胞发育中发挥重要作用。

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