论文部分内容阅读
目的探讨人胰岛素样生长因子-I(IGF-I)基因P1启动子区域单核苷酸多态性(SNP)-705T>C和-603T>A对启动子活性的影响。方法招募152名健康志愿者参与本研究,取静脉血样品,使用基因组DNA提取试剂盒从血样中提取全基因组DNA。通过限制片段长度多态性分析(RFLP),对每名志愿者的SNP-705T>C和-603T>A进行基因分型,统计各基因型的频率。使用PHASE v2.1软件程序分析P1启动子单体型的类型和频率。通过荧光素酶报告基因测定不同P1启动子单体型的活力差异。结果成功提取了每名志愿者的全基因组DNA并测定了SNP-705T>C和-603T>A的基因型。基因型-705T/T、705T/C、-705C/C的频率分别与-603T/T、-603T/A、-603A/A相同,分别为43.4%、45.4%、11.2%。等位基因-705T、-705C的频率分别与-603T、-603A相同,分别为66.1%、33.9%。SNP-705T>C和-603T>A之间存在完全的连锁不平衡,其组成的P1启动子单体型只有-705T/-603T和-705C/-603A两种,频率分别为66.1%和33.9%。报告基因分析结果表明,P1启动子-705C/-603A单体型的活力显著高于-705T/-603T单体型。结论人IGF-I基因P1启动子区域SNP-705T/C和-603T/A能够影响P1启动子的活性。
Objective To investigate the effects of single nucleotide polymorphisms (SNPs) -705T> C and -603T> A on P1 promoter region of human insulin-like growth factor-I (IGF-I) gene on promoter activity. Methods A total of 152 healthy volunteers were enrolled in this study. Venous blood samples were taken and whole genome DNA was extracted from blood samples using a genomic DNA extraction kit. By restriction fragment length polymorphism analysis (RFLP), SNP-705T> C and -603T> A for each volunteer were genotyped and the frequency of each genotype was calculated. The PHASE v2.1 software program was used to analyze the type and frequency of P1 promoter haplotypes. Differences in viability of different P1 promoter haplotypes were determined by luciferase reporter gene. Results Genomic DNA of each volunteer was successfully extracted and the genotypes of SNP-705T> C and -603T> A were determined. The frequencies of genotypes -705T / T, 705T / C and -705C / C were the same as that of -603T / T, -603T / A and -603A / A, which were 43.4%, 45.4% and 11.2% respectively. The frequencies of alleles -705T and -705C were the same as that of -603T and -603A, which were 66.1% and 33.9% respectively. There was a complete linkage disequilibrium between SNP-705T> C and -603T> A. The P1 promoter haplotype consisted of only -705T / -603T and -705C / -603A with frequencies of 66.1% and 33.9 %. Reporter gene analysis showed that the P1 promoter -705C / -603A haplotype significantly higher than the -705T / -603T haplotype. Conclusions SNP-705T / C and -603T / A of P1 promoter of human IGF-I gene can affect the activity of P1 promoter.