| 洪涛,王红卫,文格波.抑制性消减杂交法构建甲状腺癌差异表达基因文库.[J].中南医学科学杂志.,2003,(1):5-8. |
| 抑制性消减杂交法构建甲状腺癌差异表达基因文库 |
| Screening and Cloning Differentially Expressed Genes of Thyroid Carcinoma |
| 修订日期：2002-12-15 |
| DOI： |
| 中文关键词: 抑制性消减杂交法 甲状腺癌 差异表达 特异性基因 基因文库 |
| 英文关键词:thyroid neoplasms,hybridization,genes |
| 基金项目:国家自然科学基金资助项目(批准号:39970346) |
| 洪涛 王红卫 文格波 |
南华大学第一附属医院 湖南衡阳421001
(洪涛，王红卫)
，南华大学第一附属医院 湖南衡阳421001(文格波)
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| 中文摘要: |
| 目的：筛选并克隆在甲状腺癌组织与正常甲状腺组织之间差异表达的基因。方法：(1)应用抑制性消减杂交(SSH)进行基因差示表达分析，构建人甲状腺癌组织与正常甲状腺组织之间差异表达的cDNA消减文库；(2)克隆、鉴定甲状腺癌组织特异性高／低表达的基因；(3)登录Genbank，运用Blastn程序进行同源性分析。结果：成功构建了高消减效率的人甲状腺癌cDNA消减文库，对其中数个克隆的插入cDNA片段进行测序，经检索Genbank表明，正向SSH产物中有3个片段与来源于结肠上皮腺癌和Lung Epidermoid carcinoma的EST有100％同源性，提示它们来自恶性肿瘤相关基因。反向SSH产物中有5个片段为未知新序列，提示它们可能来自于在甲状腺癌中特异性低表达的新基因。结论：通过抑制性消减杂交技术，构建了人甲状腺癌cDNA消减文库，为下一步筛选鉴定甲状腺癌特异性基因及其全长克隆、功能研究等工作打下了基础。 |
| 英文摘要: |
| Objective To clone and identify differentially expressed genes which are related to thyroid carcinoma . Methods (1) Using the technique of SSH to construct a subtractive library containing differentially expressed genes' fragments. (2) Cloning and primarily analysising those differentially expressed ESTs. (3) Homology analysis of all cDNA fragments from SSH by Blastn through Genbank. Result The human TCC subtractive library with high subtractive efficiency was successfully constructed. Homology analysis of all 3 cDNA fragments from forward SSH by Blastn through Genbank showed high homology with some known "tumor related gene . On the other hand , all 5 cDNA fragments from reverse SSH showed low homology with any known gene of human beings, it' s indicated that those 5 cDNA fragments were from novel genes which specifically down - regulated expressed in TCC. Conclusion Through using SSH, we have successfully constructed the human TCC subtractive library and therefore formed a solid foundation for identifying and full - length cloning novel genes related to thyroid carcinoma cells. |
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