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目的 应用抑制性消减杂交技术 ,构建人甲状腺癌与正常甲状腺差异表达的 c DNA消减文库。方法 分别从甲状腺癌及正常甲状腺组织中提取总 RNA,应用高效、灵敏的抑制性消减杂交技术 ,构建成功 c DNA消减文库 ,再转染大肠杆菌进行文库扩增。结果 构建成功具有高消减效率的人甲状腺癌组织 c DNA消减文库 ,文库扩增得到了 4 0 0个克隆 ,随机挑取 5 0个制备质粒 ,酶切分析均得到 5 0 bp- 4 0 0 bp大小插入片段 ,这些片段可能就是载有高度特异性的目的片段。结论 人甲状腺癌组织 c DNA消减文库的建立为进一步克隆甲状腺癌特异性表达的新基因奠定了基础
Objective To construct a subtractive cDNA library of c DNA differentially expressed in human thyroid carcinoma and normal thyroid gland by suppression subtractive hybridization. Methods Total RNA was extracted from thyroid cancer and normal thyroid tissue respectively. The efficient and sensitive suppression subtractive hybridization was used to construct a successful cDNA subtractive library and then transfected into E. coli for library amplification. Results A DNA subtractive cDNA library of human thyroid carcinoma with high efficiency was established successfully. 400 clones were amplified and 50 randomly selected plasmids were obtained. The size of the insert, these fragments may be loaded with a high degree of specificity of the target fragment. Conclusion The establishment of cDNA subtractive library of human thyroid carcinoma lays the foundation for further cloning of novel genes specifically expressed in thyroid cancer