高甘油三酯血症大鼠高血凝相关基因的筛选

Screening of Hypercoagulability-related Genes in Aorta of Hypertriglyceridemic Rats

目的 筛选实验性高甘油三酯血症诱发大鼠高血凝的相关基因。方法 采用抑制性消减杂交技术构建大鼠高血凝 c DNA消减文库 ,并用半定量 RT- PCR初步鉴定差异表达基因。结果 成功构建了大鼠高血凝 c D-NA消减文库 ,2 0个阳性克隆的序列测定和同源比较表明这些克隆代表了 19个基因 ,其中 8个为已知基因但首次发现与高血凝相关 ,11个为新的 ESTs,已被 Gen Bank收录。半定量 RT- PCR证明 3个 ESTs( HC0 0 2、HC0 5 7和HC0 67)为表达上调基因。结论 所构建的消减文库消减效率高 ,为进一步筛选、克隆大鼠高血凝相关的新基因奠定了基础 ,HC0 0 2、HC0 5 7和 HC0

Objective The aim of this study was to screen hypercoagulability-related genes in aorta of hypertriglyceridemic rats. Methods Suppression subtractive hybridization (SSH) was used to construct hypercoagulable rat cDNA subtractive library. Semi-quantitative RT-PCR was applied to identify differentially expressed genes. Results Hypercoagulable rat cDNA subtractive library had been successfully constructed. Sequence analysis and homology searching of 20 clones showed that these clones represent 19 genes: 8 are known genes newly found to have a relationship to hypercoagulability and 11 are novel genes (ESTs) that had been deposited to GenBank. Three ESTs (HC002, HC057 and HC067) were confirmed to be over-expressed genes by semi-quantitative RT-PCR. Conclusion The constructed subtractive library is highly efficient and has laid a foundation for screening and cloning hypercoagulability-related novel genes of rat. HC002, HC057 and HC067 are probably related to hypercoagulability induced by hypertriglyceridemia.