中国防痨杂志2017，Vol.39Issue(8)：805-808,4.DOI:10.3969/j.issn.1000-6621.2017.08.004
功能性单核苷酸多态性调控结核病易感的分子机制研究
Molecular mechanism of functional single nucleotide polymorphisms regulated tuberculosis susceptibility
摘要
Abstract
Objective To construct different genotype dual luciferase expression plasmid expressed the rs3840880 loci of promoter region located in arachidonic acid lipoxidase-12 (ALOX-12) gene, and to analyze the effects of this single nucleotide polymorphisms (SNP) on ALOX-12 gene promoter activity and gene expression, in order to elucidate the molecular mechanism of functional SNP regulated tuberculosis susceptibility.Methods The 1700 bp fragments contained rs2840880 loci of promoter region located in ALOX-12 gene were amplified with human genome DNA extracted from whole blood samples by PCR.The pGL3-Basic-rs3840880 DEL plasmids was obtained from pGL3-Basic plasmid constructed with the PCR products and the original plasmid by site-directed mutagenesis and transfected into human cervical carcinoma cell line(Hela).The relative fluorescence values were detected by the dual luciferase assay system.Results The pGL3-Basic-DEL plasmid from pGL3-Basic-G plasmid containing the rs3840880 loci of promoter region located in the ALOX-12 gene by site-directed mutagenesis was obtained and sequenced successfully.The relative fluorescence ratio (F/R) were detected after two plasmids were transfected into Hela cell line.The fluorescence ratio of allele G (F/R=0.194) was lower than that of allele DEL (F/R=0.274), pGL3-Basic (F/R=0.129) with significant difference statistically (F=25.09,P=0.001).Conclusion The expression differences of fluorescence reporter gene in the plasmids constructed with different alleles and transfected into Hela cell line are significant statistically.It shows that functional SNP of ALOX-12 gene affect really the activity of promoter and regulate tuberculosis susceptibility.关键词
花生四烯酸盐12-脂氧合酶/多态性,单核苷酸/荧光光度测定法/结核病/基因表达调控,细菌Key words
Arachidonate 12-lipoxygenase/Polymorphism,single nucleotide/Fluorophotometry/Tuberculosis/Gene expression regulation,bacterial引用本文复制引用
朱秀云,汪文斐,张培泽,张洁云,邱智辉,王召钦,张明霞,邓国防..功能性单核苷酸多态性调控结核病易感的分子机制研究[J].中国防痨杂志,2017,39(8):805-808,4.基金项目
国家自然科学基金(81500004) （81500004）
广东省自然科学基金(2015A030313692) （2015A030313692）
深圳市科技计划项目(JCYJ20160427152106244,JCYJ20160427153348709,JCYJ20150402111430658) （JCYJ20160427152106244,JCYJ20160427153348709,JCYJ20150402111430658）
深圳市卫生计生系统科研项目(201501029) （201501029）
