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应用改良的mRNA差异展示法克隆人鼻咽上皮特异性表达的cDNA片段

Cloning of a Tissue specific cDNA Fragment from Human Nasopharyngeal Epithelium by Means of Improved mRNA Differential Display Method
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摘要 本文对mRNA差异展示法进行了改良.利用一组随机引物与来源于mRNA翻译起始位点区域的引物组合进行RT-PCR,使差异展示的片段来源于蛋白编码区,并对差异展示的条件进行了优化.应用此法对人鼻咽上皮与软腭口腔粘膜上皮的基因表达进行了比较研究,得到了10个在鼻咽上皮特异表达的cDNA片段,并对其中的5个片段进行了亚克隆和序列分析.通过与GeneBank数据库中的序列进行同源性比较,确定其中两个片段为未知新序列,Northern杂交证实其中一个片段NES1为鼻咽上皮特异性表达片段. The method of mRNA differential display was improved by using a set of arbitrary primers combined with primers from the translation start site.In this case,the cDNA fragments of differential display originated from protein coding region of mRNA.Selecting different RT PCR reaction conditions and different recovery method of fragments optimized differential display.Gene expression profiles of human nasopharyngeal epithelium was compared with that of human oral cavity epithelium of soft palate by this method,and it were found that ten cDNA fragments uniquely expressed in nasopharyngeal epithelium. Five of them were subcloned and sequenced,and two were found to be novel sequences byBLAST sequence similarity searching in GeneBank nucleotide sequence database.Finally,one of them,NES1,was confirmed to be a tissue specific cDNA fragment of human nasopharyngeal epithelium.
出处 《生命科学研究》 CAS CSCD 1998年第3期182-188,共7页 Life Science Research
基金 国家自然科学基金 美国中华医学基金会基金
关键词 mRNA差异展示法 人鼻咽上皮 亚克隆 组织特异性表达 mRNA differential display method,human nasopharyngeal epithelium,cloning,tissue specific expression
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参考文献7

  • 1Yao KT,Li ZK. Development and differentiation of human fetal nasopbsryngeal epithelium. Abstract of 3rd Hong Kong International Cancer Congress & 7th International EBV Symposium Hong Kong, 1996-175.
  • 2Liang P, Pardee AB. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science, 1992,257:967- 971.
  • 3Liang P,Averbou KH,Aeyomoorsi K et al. Differential display and cloning of messenger RNAs from human breast cancer versus mammary epithelial ceils. Cancer Res. 1992,52 :6966 - 6968.
  • 4Sun Y,Glenn H,Colburn NH. Molecular cloning of five messenger RNAs differentially expressed in preneoplastic or neoplastic JB5 mouse epidermal cells :One is homologous to human tissue inhibitor of metalloproteinase-3. Cancer Res. 1994,54 : 1139- 1144.
  • 5Kozak M. An analysis of vertebrate mRNA sequences:intimations of translational control, J Cell Biol.1991, 115 : 887- 903.
  • 6Lisitsyn NA,Lisitsyn NM,Wigler M et al. Cloning the differences between two complex genomes. Science, 1993,259:946-951.
  • 7Diatcbenko L,Chris LYF,Campbell AP et al. Suppression subtractive bybridization:a method for generating differentially regulated or tissue-specific cDNA probes and libraries. Pro Natl Acad Set USA.93:6025-6030.

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