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目的 :杰出运动能力受控于基因与环境的相互作用 ,由多基因控制。寡核苷酸芯片技术是近年出现的一类基因结构分析技术 ,此项技术出现使基因突变检测更加程序化和规模化。本研究选取与运动能力有关的胰岛素生长因子Ⅱ (IGF -Ⅱ )基因为该方法学及其应用研究的突破点 ,尝试把DNA芯片技术引入体育科研。方法 :寡核苷酸芯片技术。结果 :(1)IGF -Ⅱ基因SNP的分析中 ,基因组DNA采用 1∶2 0的套式扩增 ,杂交液中加氯化四甲氨 ,杂交温度 4 2℃ ,杂交时间 6 0min效果最好 ;(2 )国家自行车队的 6名队员基因型相同。结果表明 :(1)PCR产物可以直接用于检测突变 ,不对称PCR产物杂交阳性点信号更强 ;(2 )由碱基组成计算的Tm值与其实际的Tm值有一定的差距 ,会影响杂交温度 ;(3)有效的DNA载波片的处理效果直接影响杂交结果 ,是芯片制备中关键环节之一 ;(4)IGF -Ⅱ是研究运动能力相关基因的一个很好的候选基因。
Purpose: Outstanding motor ability is controlled by the interaction of genes with the environment and controlled by multiple genes. Oligonucleotide microarray technology is a kind of gene structure analysis technology that appears in recent years. The technology appears to make gene mutation detection more program and scale. In this study, we selected the insulin-like growth factor-Ⅱ (IGF-Ⅱ) gene related to exercise capacity as a breakthrough point of the methodology and its application, and tried to introduce DNA chip technology into the research of sports science. Methods: Oligonucleotide chip technology. Results: (1) In the analysis of SNP of IGF-Ⅱ gene, the genomic DNA was amplified by 1: 200 nested PCR. Tetramethylammonium chloride was added to the hybridization solution at a temperature of 42 ℃ and the hybridization time was 60 min ; (2) 6 members of the National Cycling Team have the same genotype. The results showed that: (1) PCR products could be directly used to detect mutations, and asymmetric PCR products showed stronger signal of positive hybridization; (2) Tm values calculated from base composition had a certain gap with their actual Tm values, which would affect the hybridization Temperature; (3) The effective DNA carrier processing effect directly affects the hybridization results, is one of the key aspects in chip preparation; (4) IGF-Ⅱ is a good candidate gene for the study of exercise-related genes.