目的利用DNA-pool技术对原发性高血压患者进行测序,探索中国原发性高血压患者单核苷酸多态性变异(SNP)情况。方法连续收集2014年3月至2014年6月深圳市孙逸仙心血管医院的高血压门诊患者100例。将基因组DNA片段化处理至400~800bp进行建...目的利用DNA-pool技术对原发性高血压患者进行测序,探索中国原发性高血压患者单核苷酸多态性变异(SNP)情况。方法连续收集2014年3月至2014年6月深圳市孙逸仙心血管医院的高血压门诊患者100例。将基因组DNA片段化处理至400~800bp进行建库测序,将测序结果与NCBI(National Center of Biotechnology Information)人类基因库hg19进行比对。结果共生成120.8Gb原始序列数据,测序深度约为36.13倍,覆盖率达到了99.88%。通过生物信息学分析共检测到4 305 668个SNP点,其中C:G→T:A的变异类型最多,达到12 314个变异位点。结论研究验证了使用DNA-pool的全基因组重排序的方法。研究所得数据也对中国原发性高血压的基因型数据库进行了补充,对未来原发性高血压基因研究提供了一定的帮助。展开更多
Objective:To investigate the effects of -2242,-1892 and -1837 single nucleotide polymorphisms(SNPs) on toll-like receptor 4(TLR4) promoter activity.Methods:Polymerase chain reaction(PCR) and site direct mutation techn...Objective:To investigate the effects of -2242,-1892 and -1837 single nucleotide polymorphisms(SNPs) on toll-like receptor 4(TLR4) promoter activity.Methods:Polymerase chain reaction(PCR) and site direct mutation technology were used to construct TLR4 basic promoter and -2242C,-1892A and -1837G mutate promoter plasmids.Dual-Luciferase Reporter assay system was used to detect the activity of constructed promoter following human embryonic kidney(HEK) 293 cells were transiently cotransfected with the constructed plasmids and the control plasmid pRL-CMV.Results:In HEK293 cells,the activity of -2242C mutate promoter was higher than -2242T promoter,and there was no significant difference when both -1892A and -1837G mutate promoter compared with -1892G and -1837A promoter,respectively.Conclusion:It is implied that -2242T→C base variation can enhance the activity of TLR4 promoter,while -1892 and -1837 SNPs have no effect on TLR4 promoter activity.展开更多
文摘目的利用DNA-pool技术对原发性高血压患者进行测序,探索中国原发性高血压患者单核苷酸多态性变异(SNP)情况。方法连续收集2014年3月至2014年6月深圳市孙逸仙心血管医院的高血压门诊患者100例。将基因组DNA片段化处理至400~800bp进行建库测序,将测序结果与NCBI(National Center of Biotechnology Information)人类基因库hg19进行比对。结果共生成120.8Gb原始序列数据,测序深度约为36.13倍,覆盖率达到了99.88%。通过生物信息学分析共检测到4 305 668个SNP点,其中C:G→T:A的变异类型最多,达到12 314个变异位点。结论研究验证了使用DNA-pool的全基因组重排序的方法。研究所得数据也对中国原发性高血压的基因型数据库进行了补充,对未来原发性高血压基因研究提供了一定的帮助。
基金Supported by the Major State Basic Research Development Program of China (2005CB522602)the National Funds for Outstanding Youth Scientists (30325040)
文摘Objective:To investigate the effects of -2242,-1892 and -1837 single nucleotide polymorphisms(SNPs) on toll-like receptor 4(TLR4) promoter activity.Methods:Polymerase chain reaction(PCR) and site direct mutation technology were used to construct TLR4 basic promoter and -2242C,-1892A and -1837G mutate promoter plasmids.Dual-Luciferase Reporter assay system was used to detect the activity of constructed promoter following human embryonic kidney(HEK) 293 cells were transiently cotransfected with the constructed plasmids and the control plasmid pRL-CMV.Results:In HEK293 cells,the activity of -2242C mutate promoter was higher than -2242T promoter,and there was no significant difference when both -1892A and -1837G mutate promoter compared with -1892G and -1837A promoter,respectively.Conclusion:It is implied that -2242T→C base variation can enhance the activity of TLR4 promoter,while -1892 and -1837 SNPs have no effect on TLR4 promoter activity.
