[1]杨俊红),崔新征),方华),等.重症肌无力患者胸腺组织抑制性消减cDNA文库的构建及分析*[J].郑州大学学报(医学版),2012,(03):295.
 YANG Junhong),CUI Xinzheng),FANG Hua),et al.Construction and analysis of suppression subtractive cDNA library of thymus from myasthenia gravis patients[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2012,(03):295.
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重症肌无力患者胸腺组织抑制性消减cDNA文库的构建及分析*
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2012年03期
页码:
295
栏目:
专题研究
出版日期:
2012-05-20

文章信息/Info

Title:
Construction and analysis of suppression subtractive cDNA library of thymus from myasthenia gravis patients
作者:
杨俊红1)崔新征12)方华4)关兵峰12)赵国强5)张清勇12)#高峰4)
1)河南中医学院第一附属医院脑病1区 郑州 450003 2)郑州大学第二附属医院胸外科 郑州 450014 3)郑州大学生物医学工程研究所 郑州 450014 4) 郑州大学医药科学研究院免疫学研究室神经免疫实验室 郑州 450052 5)郑州大学基础医学院微生物学与免疫学教研室 郑州 450001
Author(s):
YANG Junhong1) CUI Xinzheng12)FANG Hua4)GUAN Bingfeng12)ZHAO Guoqiang5)ZHANG Qingyong12)GAO Feng4)
1)Department of Encephalopathy Zone 1,the First Affiliated Hospital,Henan University of Traditional Chinese Medicine,Zhengzhou 450003 2)Department of Thoracic Surgery, the Second Affiliated Hospital, Zhengzhou University, Zhengzhou 450014 3)Institute of Biological Medicine,Zhengzhou University, Zhengzhou 450014
关键词:
抑制性消减重症肌无力差异基因cDNA文库胸腺
Keywords:
suppression subtractive myasthenia gravis differential genecDNA librarythymus
分类号:
R593.2
摘要:
目的:构建重症肌无力(MG)患者胸腺组织抑制性消减cDNA文库,分析MG胸腺差异表达基因。方法:分别从6例正常和6例MG患者胸腺组织中分离出mRNA,逆转录合成cDNA,行抑制性消减杂交,将得到的差异表达基因进行TA克隆,构建cDNA文库,并进行序列分析。应用实时荧光定量PCR检测20例MG患者、10例正常对照胸腺组织中GSTM3和KPNA5 mRNA的表达。结果:共获得125个阳性克隆;Blast同源性检索共得到27个差异表达基因;MG患者GSTM3、KPNA5基因表达量(0.671±0.097和0.712±0.080)高于正常对照胸腺(0.582±0.047和0.571±0.018),差异有统计学意义(tGSTM3=5.458,P<0.001;tKPNA5=2.755,P=0.010),与消减文库结果一致。结论:成功建立了MG胸腺组织抑制性消减cDNA文库。
Abstract:
Aim:To build suppression subtractive cDNA library of thymus from myasthenia gravis (MG) patients.Methods:The total mRNA was extracted from thymus of normal people and MG patients and synthethized to cDNA by reverse transcription.The differentially expressed genes obtained by using suppression subtractive hybridization were proceeded by TA cloning and sequence analysis.The expressions of GSTM3 and KPNA5 in thymus of 20 MG patients and 10 normal people were detected by realtime quantitative PCR.Results:A total of 125 positive clones were obtained;27 differentially expressed genes were obtained by Blast homology search.The expression levels of GSTM3, KPNA5 in thymus tissue of MG patients(0.671±0.097,0.712±0.080)were significantly higher than those of the control group (0.582±0.047,0.571±0.018)(tGSTM3=5.458,P<0.001;tKPNA5=2.755,P=0.010),consistent with the result of subtractive library.Conclusion:The establishment of suppression subtractive cDNA library of MG thymus is successful.

参考文献/References:

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备注/Memo

备注/Memo:
*河南省杰出青年基金资助项目0212000100;河南省科研院所专项基金资助项目0441130303 #通讯作者,男,1963年11月生,主任医师,教授,研究方向:重症肌无力的临床医学与发病机制,Email:qyzhang@zzu.edu.cn
更新日期/Last Update: 2012-05-31