Zhao Yuhua, Liu Bingwen, Bai Huai
Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2003 Jul;34(3):401-4.
The aim of this study was to screen hypercoagulability-related genes in aorta of hypertriglyceridemic rats.
Suppression subtractive hybridization (SSH) was used to construct hypercoagulable rat cDNA subtractive library. Semi-quantitative RT-PCR was applied to identify differentially expressed genes.
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.
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.
本研究旨在筛选高甘油三酯血症大鼠主动脉中与高凝性相关的基因。
采用抑制性消减杂交（SSH）构建高凝大鼠cDNA消减文库。应用半定量RT-PCR鉴定差异表达基因。
成功构建了高凝大鼠cDNA消减文库。对20个克隆进行序列分析和同源性搜索表明，这些克隆代表19个基因：8个是新发现与高凝性有关的已知基因，11个是已提交到GenBank的新基因（ESTs）。通过半定量RT-PCR证实3个ESTs（HC002、HC057和HC067）是高表达基因。
构建的消减文库效率高，为大鼠高凝性相关新基因的筛选和克隆奠定了基础，HC002、HC057和HC067可能与高甘油三酯血症诱导的高凝性有关。
