快速老化模型小鼠海马正反向抑制消减cDNA文库的构建

目的:构建快速老化模型小鼠(SAM)海马正反向抑制消减cDNA文库,以揭示SAMP8学习记忆脑老化的机制,同时为研究阿尔茨海默病(AD)的发病机制提供线索。方法:以快速老化模型小鼠SAMP8和SAMR1海马的总RNA为材料,采用抑制消减杂交方法和蓝白斑筛选克隆构建文库,并用PCR鉴定了文库的质量。结果:成功构建了12月龄雄性SAMP8和SAMR1海马的正反向抑制消减cDNA文库,其中正向文库包含864个克隆,反向文库包含960个克隆,阳性克隆率为96.16%,插入片段范围为250～2000bp。结论:SAMP8和SAMR1海马的正反向抑制消减cDNA文库的构建,为进一步筛选鉴定SAMR1和SAMP8海马差异表达基因提供了丰富的实验材料。

Construction of the Forward and Reverse Suppression Subtracted cDNA Library of Hippocampus of Senescence-Accelerated Mouse

Objective: In order to disclose the mechanism of SAMP8's brain aging and provide some data for the pathogenesis of Alzheimer's disease(AD),the forward and reverse suppression subtracted cDNA library of hippocampus of senescence-accelerated mouse(SAM) were constructed.Methods: Suppression subtractive hybridization(SSH) technique was used to construct the subtracted hippocampal cDNA library of SAMP8 and SAMR1,and the subtracted cDNA library was screened by blue/white blot and analysised by PCR.Results: The forward library contained 864 clones and 960 clones in the reverse library,the positive ratio of constructed cDNA libraries was 96.16% and the length of cDNA fragments was ranged from 250 to 2 000 bp.Conclusion: The forward and reverse suppression subtracted cDNA library of hippocampus of SAM was successfully constructed.The two libraries would become the abundance material used to screen and decide the hippocampal differential expression genes of SAMP8 and SAMR1.