快速老化小鼠海马抑制消减cDNA文库的构建

Construction of suppression subtracted cDNA library of hippocampus of senescence-accelerated mouse

目的 :构建快速老化小鼠 (senescence_acceleratedmouse ,SAM)海马抑制消减cDNA文库。方法 :以SAM快速老化亚系SAM_prone 8(SAMP8)海马作为实验组 ,抗快速老化小鼠亚系SAM_resistance 1(SAMR1)海马作为对照组 ,应用抑制消减杂交 (SSH)技术 ,构建SAMP8海马特异表达cDNA抑制消减文库 ;用蓝白斑筛选 ,以PCR鉴定文库质量。结果 :构建文库的阳性克隆率为 96 .18% ,克隆中cDNA片段介于 2 5 0～ 2 0 0 0bp之间。结论 :成功构建了SAMP8海马抑制消减文库。

Objective: To construct the suppression subtracted library of hippocampus of senescence-accelerated mouse (SAM). Methods: Suppression subtractive hybridization (SSH) technique was used to construct the subtracted hippocampal cDNA library of SAM. The hippocampal cDNA of SAMP8 (SAM-prone/8), a senescence-accelerated substrain of SAM,was used as tester, while that of SAMR1 (SAM-resistance/1), a senescence-resistant substrain of SAM,as driver. The subtracted cDNA library was screened by blue/white blot and analysed by PCR. Results: The positive ratio of constructed cDNA library was 96.18% and the length of cDNA fragments ranged from 250 to 2?000?bp. Conclusion: The suppression subtracted cDNA library of hippocampus of SAM is successfully constructed.