目的:探讨健康中国汉族人群中basigin(BSG)基因的单核苷酸多态性(single nucleotide polymorphisms,SNPs)发生情况。方法:随机收集48例健康、无亲缘关系的中国汉族人外周血液并提取基因组DNA,设计引物对所有个体BSG基因的启动子区、外显子区和外显子内含子交界区的序列进行PCR扩增和正反向测序,通过判读测序峰图,明确SNPs的发生情况及其频率;通过Hardy-Weinberg平衡分析、单倍型推测和连锁不平衡分析,确定BSG基因位点的单倍型标签SNPs(haplotype tag SNPs,htSNPs);中性理论检验查明该基因位点SNPs频率分布是否符合选择中性。结果:共发现19个SNPs,其中包括2个新发现的SNPs;所有SNPs位点基因型分布均符合Hardy-Weinberg平衡。该基因位点共推测出4种常见单倍型域,确定9个SNPs为htSNPs。中性理论检验结果提示健康中国汉族人群BSG基因的SNPs分布符合中性进化假说。结论:首次对中国健康汉族人群BSG基因的SNPs进行了发掘,确定了其9个单倍型标签SNPs,为在汉族人群中研究该基因的遗传多态性与疾病易感性或药物反应性个体差异奠定了基础。
OBJECTIVE To investigate whether micro RNA(mi RNA)mi R-21 regulates dimethylarginine dimethylaminohydrolase 1(DDAH1)expression through binding 3′-UTR regiondirectly in human umbilical venous endothelial cells(HUVECs)and to explore whether DDAH1-V2/V3 transcripts can function as micro RNA sponge,thereby modulating DDAH1-V1 expression.METHODS The DDAH13′-UTR containing mi R-21 recognizing sequence was cloned into Pmir GLO dual-luciferase mi RNA target expression plasmid to construct PmirGLO-mi R-21.The plasmid and mi R-21(at concentrations of 25,50,100 nmol·L-1,respectively)or negative control(100 nmol·L-1)were co-transfected into HUVECs,luciferase activity was detected at 24 h.HUVECs were incubated with 2μg·m L-1actinomycin D for the indicated time after mi R-21(25 nmol.L-1)transfection,half-lives of DDAH1 m RNA were determined.HUVECs were transfected with Pmir GLO-mi R-21 alone or co-transfected with mi R-21 for 24 h,DDAH1 transcripts m RNA and DDAH1protein expression were determined.RESULTS Mi R-21decreased luciferase activity of Pmir GLO-mi R-21 in a dose-dependent manner(P<0.05 for 25 nmol·L-1mi R-21,P<0.01 for 50 nmol·L-1and 100 nmol·L-1mi R-21),and mi R-21 inhibitor increased reporter activity of PmirGLO-mi R-21 and m RNA expression of all DDAH1 three transcript variants significantly(P<0.05,respectively).The degree of increase in endogenous DDAH1 m RNA expression by mi R-21 inhibitor was more obvious for DDAH1-V3.Overexpression of mi R-21 decreased m RNA expression and m RNA half-life time of all DDAH1 transcripts significantly(P<0.05),and DDAH1-V2 displayed significantly decreased half-life time than DDAH1-V1and-V3 with or without mi R-21 transfection(P<0.05,respectively).Mi R-21(100 nmol·L-1)decreased DDAH1protein expression significantly(P<0.05),which was reversed by Pmir GLO-mi R-21 transfection(P<0.05).Transfection of Pmir GLO-mi R-21 alone increased intracellular mi R-21 expression by approximately 5.6-fold,but only showed a trend of increase in DDAH1 protein expression.CONCLUSION Our resu
