[小鼠胸腺衰老相关基因的差异表达分析与克隆]

[Differential expression analysis and cloning of murine thymic aged-related genes].

作者信息

Li Y H, Su H, He W, Cui L X, Hiromoto O

机构信息

Department of Immunology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.

出版信息

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2001 Apr;23(2):158-62.

PMID:12905894

Abstract

OBJECTIVE

Differential expression analysis and cloning of murine thymic aged-related genes.

METHODS

Different expressions of thymic mRNAs from 1- and 10-month old mice were analyzed via DDRT-PCR and different expression sequence tags (ESTs) were obtained, following by identification with Northern Blotting, DNA sequencing, as well as screening of cDNA library.

RESULTS

It was found that there would be a significant difference of gene expression in murine thymuses. Gene expression of some genes were exclusive in thymus from 1 or 10-month old mice, while some expressed different with age. 108 differential display cDNA fragments were recovered, among which 31 were positive for hybridization by Northern blot. After sequenced, 14 ESTs were found to share high homology to known genes, whereas remaining 17 were novel. A murine thymic cDNA library was screened by using one cDNA fragment that expressed with higher level in total RNA of 1-month old murine thymic tissues than 10-month old. Finally, one 1,470 bp fragment was cloned and showed a 99% of homology to murine transketolase.

CONCLUSION

Expression of murine thymic genes has displayed a marked difference with age. These genes might be participated in thymic atrophy.

摘要

目的

对小鼠胸腺衰老相关基因进行差异表达分析及克隆。

方法

采用DDRT-PCR分析1月龄和10月龄小鼠胸腺mRNA的差异表达，获得差异表达序列标签（ESTs），随后通过Northern印迹、DNA测序以及cDNA文库筛选进行鉴定。

结果

发现小鼠胸腺中基因表达存在显著差异。一些基因的表达在1月龄或10月龄小鼠的胸腺中是特异性的，而一些基因的表达随年龄不同而变化。回收了108个差异显示cDNA片段，其中31个经Northern印迹杂交呈阳性。测序后，发现14个ESTs与已知基因具有高度同源性，其余17个为新基因。用一个在1月龄小鼠胸腺组织总RNA中表达水平高于10月龄的cDNA片段筛选小鼠胸腺cDNA文库。最终，克隆到一个1470 bp的片段，与小鼠转酮醇酶具有99%的同源性。