中国医学科学院学报
中國醫學科學院學報
중국의학과학원학보
ACTA ACADEMIAE MEDICINAE SINICAE
2001年
2期
158-162
,共5页
李永海%苏华%何维%崔莲仙%越智宏伦
李永海%囌華%何維%崔蓮仙%越智宏倫
리영해%소화%하유%최련선%월지굉륜
DDRT-PCR小鼠胸腺基因差异表达
DDRT-PCR小鼠胸腺基因差異錶達
DDRT-PCR소서흉선기인차이표체
目的小鼠胸腺增龄相关性基因差异表达研究及其克隆。方法利用DDRT-PCR技术对1月龄和10月龄小鼠胸腺mRNA差异表达进行分析，获得差异表达片段(ESTs)。进一步作Northern印迹分析并进行核苷酸测序。选取其中一个EST为探针，从小鼠胸腺cDNA文库中克隆基因。结果对1月龄和10月龄小鼠胸腺mRNA差异表达进行DDRT-PCR分析，发现小鼠胸腺基因存在增龄相关性的表达差异，某些基因在1或10月龄胸腺内有选择性表达，某些基因仅有表达量的变化。共获得108个差异表达的ESTs，其中31个呈Northern印迹阳性，全部测序。经同源性分析，有14个ESTs与已知基因有较高的同源性，17个ESTs为新的cDNA片段。选取其中1个在1月龄鼠胸腺高水平表达的EST，从小鼠胸腺cDNA文库中克隆到1个与小鼠的LAF1转酮醇酶高度同源的1470bp片段。结论小鼠胸腺内存在增龄性基因表达差异。据此差异所获得的胸腺差异表达基因可能参与胸腺衰老与萎缩过程。
目的小鼠胸腺增齡相關性基因差異錶達研究及其剋隆。方法利用DDRT-PCR技術對1月齡和10月齡小鼠胸腺mRNA差異錶達進行分析，穫得差異錶達片段(ESTs)。進一步作Northern印跡分析併進行覈苷痠測序。選取其中一箇EST為探針，從小鼠胸腺cDNA文庫中剋隆基因。結果對1月齡和10月齡小鼠胸腺mRNA差異錶達進行DDRT-PCR分析，髮現小鼠胸腺基因存在增齡相關性的錶達差異，某些基因在1或10月齡胸腺內有選擇性錶達，某些基因僅有錶達量的變化。共穫得108箇差異錶達的ESTs，其中31箇呈Northern印跡暘性，全部測序。經同源性分析，有14箇ESTs與已知基因有較高的同源性，17箇ESTs為新的cDNA片段。選取其中1箇在1月齡鼠胸腺高水平錶達的EST，從小鼠胸腺cDNA文庫中剋隆到1箇與小鼠的LAF1轉酮醇酶高度同源的1470bp片段。結論小鼠胸腺內存在增齡性基因錶達差異。據此差異所穫得的胸腺差異錶達基因可能參與胸腺衰老與萎縮過程。
목적소서흉선증령상관성기인차이표체연구급기극륭。방법이용DDRT-PCR기술대1월령화10월령소서흉선mRNA차이표체진행분석，획득차이표체편단(ESTs)。진일보작Northern인적분석병진행핵감산측서。선취기중일개EST위탐침，종소서흉선cDNA문고중극륭기인。결과대1월령화10월령소서흉선mRNA차이표체진행DDRT-PCR분석，발현소서흉선기인존재증령상관성적표체차이，모사기인재1혹10월령흉선내유선택성표체，모사기인부유표체량적변화。공획득108개차이표체적ESTs，기중31개정Northern인적양성，전부측서。경동원성분석，유14개ESTs여이지기인유교고적동원성，17개ESTs위신적cDNA편단。선취기중1개재1월령서흉선고수평표체적EST，종소서흉선cDNA문고중극륭도1개여소서적LAF1전동순매고도동원적1470bp편단。결론소서흉선내존재증령성기인표체차이。거차차이소획득적흉선차이표체기인가능삼여흉선쇠로여위축과정。
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 l-month old murine thymic tissues than 10-montholds. Finally, one 1470 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.
