【摘要】 目的 :构建不稳定型心绞痛淋巴细胞差异表达cDNA消减文库。方法 :将不稳定型心绞痛病人 (n =15 )和稳定型心绞痛病人 (n =15 )的淋巴细胞的RNA进行抑制性消减杂交 (SSH) ,将获得的顺向和逆向消减产物分别与T/A载体连接 ,采用 1×TSS一步法转化大肠杆菌JM10 9,获得消减cDNA文库 ,采用蓝白斑筛选和菌落PCR筛选有插入片段的克隆。结果 :各组cDNA文库各获得 2 0 0 0个阳性克隆 ,cDNA片段大部分分布于 ( 2 0 0 - 6 0 0 )bp之间。结论 :本研究构建了不稳定型心绞痛淋巴细胞消减cDNA文库 ,此cDNA文库有助于进一步筛选不稳定型心绞痛淋巴细胞中的差异表达基因更多还原

【Abstract】 AIM: To construct a subtracted cDNA library of differentially expressed genes in human unstable angina lymphocytes. METHODS: Suppression subtractive hybridizations (SSH) were performed between the patients with unstable angina pectoris and stable angina pectoris. Lymphocyte RNA, the obtained forward and reverse cDNA fragments were directly inserted into T/A cloning vector and transformed into E.coli JM 109 to construct a subtractive cDNA library. The inserting fragments were screened by blue and while blot screening and bacterium liqulid PCR. RESULTS: Each subtractive cDNA library contained more than 2000 positive bacteria clones. Most of them distributed between 200-600 bp inserts. CONCLUSION: The library is efficient and lays solid foundation for screening and cloning new and specific expressed genes in unstable angina lymphocyte RNA.更多还原