中文摘要：

目的：制备大鼠早期生长反应基因（Egr）-1分子探针,为进一步应用其研究Egr-1在病变组织的表达奠定基础。方法：提取大鼠心肌组织总RNA,PCR扩增基因片段,克隆入pGEM-T Easy载体,行酶切和基因测序。用随机引物DNA标记法制备α-32P标记的Egr-1和β-actin探针。Northern blot和Western blot法分析验证探针的信号强度及特异性。结果：PCR扩增所得目的基因片段大小与理论预期一致,测序证实获得基因序列与实验设计吻合。Northern blot显示该探针可检测心肌组织中目标mRNA水平,Western blot法检测到大鼠心肌组织中Egr-1蛋白的存在。结论：本研究成功制备大鼠Egr-1 cDNA探针,可用于进一步研究Egr-1与相关心血管疾病病理机制的关系。

英文摘要：

Objective：To prepare the DNA probe of rat early growth response gene（Egr）-1 for mRNA detection in injured tissues. Method：Total RNA extracted from rat myocardial tissue. Target genes were amplified by PCR and cloned into pGEM-T Easy vectors. The sequences were confirmed by enzymes digestion and DNA sequencing. The 32P-lableled rat Egr-1 cDNA probe was generated with the Random Primers DNA Labeling System. Signal intensity and specificity of the probes were verified by Northern blot analysis and further validated by Western blot analysis. Results：The PCR product matched the expected size. DNA sequencing showed a high nucleotide homology with the theoretical sequence. Northern blot indicated that these probes were applicable to determine the Egr-1 mRNA levels in myocardial tissue,which was supported by Western blot analysis. Conclusion：The successfully obtained probes in the present study will facilitate us to further study the role of Egr-1 in the pathogenesis of relevant cardiovascular disease.