儿童STING基因启动子区序列多态性分析
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国家重点研发项目(W2016EWJS03)


Sequence polymorphism of the promoter region of STING gene in children
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    摘要:

    目的:分析儿童外周血干扰素基因刺激因子(stimulator of interferon genes,STING)启动子-436~+267区域序列变化。方法:采用PCR产物直接测序的方法,对48例儿童的STING基因启动子区进行序列分析,构建重组报告质粒pGL-3/STING(TT)和pGL-3/STING(CC),转染人类胚胎肾(human embryonic kidney,HEK)293T细胞,检测其荧光素酶活性,计算相对活性单位(the relative luciferase activity unit,RLU)。结果:发现STING基因启动子区的1个新的单核苷酸多态性(single nucleotide polymorphism,SNP)位点(-401)T/C;该位点TT、TC、CC这3种基因型的频率分布分别为45.8%、37.5%、16.7%;经酶切、测序鉴定,成功构建了含有该位点的STING启动子序列(-436~+267)的表达质粒pGL-3/STING(TT)和pGL-3/STING(CC)。与pGL-3/STING(TT)质粒比较,pGL-3/STING(CC)启动子的RLU明显降低。结论:STING基因(-401)T/C是一个新的SNP,该SNP可降低STING基因启动子的活性,进而可能影响基因的转录调控,影响机体对疾病的易感性。

    Abstract:

    Objective:To explore the sequence variation of the promoter region(-436~+267)on stimulator of interferon genes(STING)of children. Methods:The sequences of the promoter region of 48 children were amplified by PCR and then sequenced. The luciferase reporter plasmid pGL-3/STING(TT)and pGL-3/STING(CC)were constructed to transfect human embryonic kidney(HEK)-293T cells, and then the luciferase gene expression was detected and the relative luciferase activity unit(RLU)was calculated. Results:A new single nucleotide polymorphism(SNP)T/C(-401)in STING promoter region was identified. The frequencies of genotype TT,TC,and CC were 45.8%,37.5%,and 16.7% respectively. Restriction endonuclease analysis and DNA sequencing verified the successful construction of the plasmid pGL-3/STING(TT)and pGL-3/STING(CC)including the SNP. Compared with plasmid pGL-3/STING(TT),pGL-3/STING(CC)exhibited a weaker promoter activity. Conclusion:T/C(-401)in STING promoter region is a new SNP. The genotype of this new SNP may reduce the activity of STING promoter,and consequently affect the transcription and expression of STING gene,and affect the body’s susceptibility to disease.

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王艳艳,郦银芳,张 莉,王晓花.儿童STING基因启动子区序列多态性分析[J].南京医科大学学报(自然科学版),2019,(10):1449-1452

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  • 收稿日期:2019-07-17
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  • 在线发布日期: 2019-11-01
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