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目的 :应用cDNAchip(cDNA芯片或微矩阵基因芯片 )技术筛选运动性心肌肥大相关基因。方法 :摘取运动组和对照组小鼠心脏 ,按一步法抽提总RNA并纯化mRNA ,将 2 30 4个点包括10 3个阴性及对照基因和 2 2 0 1条小鼠靶基因的PCR扩增产物 ,按微矩阵 (12× 12点× 16亚矩阵 ,点间距离 375 μm)点制于硅烷化玻片上 ,制备成cDNA芯片。将等量抽提纯化的运动组和对照组小鼠心肌组织mRNA ,分别与掺入的荧光分子Cy3和Cy5经逆转录合成cDNA荧光探针 ,混合后再与cD NA芯片杂交 ,通过芯片扫描仪对荧光信号图像进行扫描 ,经计算机分析比较运动组和安静对照组心肌组织中基因表达谱的变化。结果 :在 2 2 0 1条待研究基因中 ,两组小鼠心肌组织间存在差异表达的基因。具有显著表达差异的基因有 71条 ,其中上调基因有 37条 ,下调基因有 34条。结果显示 :cDNA芯片技术筛选运动性心肌肥大相关基因具有高通量、高敏度、高效率、大规模和并行性等优点。
Objective: To screen genes related to exercise-induced cardiac hypertrophy by cDNAchip (cDNA microarray or microarray). Methods: The hearts of mice in both exercise and control groups were harvested and the total RNA was extracted and purified by one-step method. PCR was performed on 2 304 samples including 10 3 negative and control genes and 2020 mouse target genes The amplified product was spotted on a silanized glass slide using a micro-matrix (12 × 12 dots × 16 sub-matrix with a distance of 375 μm) to prepare a cDNA chip. The same amount of extracted purified myocardial tissue mRNA and control group, respectively, with the incorporation of fluorescent molecules Cy3 and Cy5 by reverse transcription of cDNA fluorescent probe, mixed with cD NA chip hybridization, chip scanner Fluorescent signal images were scanned and the changes of gene expression profiles in myocardial tissue of exercise group and quiet control group were analyzed by computer. Results: Among the 2220 genes to be studied, there were differentially expressed genes between the two groups of mouse myocardium. There were 71 genes with significant difference, of which 37 genes were up-regulated and 34 genes were down-regulated. The results showed that the cDNA microarray technology has the advantages of high throughput, high sensitivity, high efficiency, large-scale and parallelism in screening genes related to exercise-induced cardiac hypertrophy.