南京医科大学学报（英文版）
南京醫科大學學報（英文版）
남경의과대학학보（영문판）
JOURNAL OF NANJING MEDICAL UNIVERSITY(ENGLISH EDITION)
2003年
6期
277-282
,共6页
肾上腺素受体α2A%单核苷酸多态性%基因表达
腎上腺素受體α2A%單覈苷痠多態性%基因錶達
신상선소수체α2A%단핵감산다태성%기인표체
α2A-adrenergic receptor%single nucleotide polymorphism%gene expression
目的:在中国人群肾上腺素α2A受体(α2A-AR)基因中搜寻单核苷酸多态性(SNP)并研究其对基因表达的影响.方法:对α2A-AR基因进行序列分析,搜寻SNP.从全血中提取基因组DNA,扩增包含G/C多态性位点的239 bp DNA片段.利用PCR-RFLP方法对中国北方人群α2A-AR基因-1296 bp位点SNP进行基因分型.利用凝胶阻滞方法研究含G/C的390 bp DNA片段与细胞核提取物的结合.结果:在中国北方人群α2A-AR基因中共发现2处SNP.-1296位SNP等位基因G、C频率分别为0.61和0.39,基因型GG,GC,CC频率分别为0.34,0.54,0.13.EMSA结果表明,细胞核提取物与-1 296 bp为C的DNA片段可特异性结合.结论:中国北方人群α2A-AR基因中存在2处SNP,调控区(-1 296位)SNP等位基因为C的DNA片段可与特异性蛋白质结合,可能通过这一机制影响基因表达.
目的:在中國人群腎上腺素α2A受體(α2A-AR)基因中搜尋單覈苷痠多態性(SNP)併研究其對基因錶達的影響.方法:對α2A-AR基因進行序列分析,搜尋SNP.從全血中提取基因組DNA,擴增包含G/C多態性位點的239 bp DNA片段.利用PCR-RFLP方法對中國北方人群α2A-AR基因-1296 bp位點SNP進行基因分型.利用凝膠阻滯方法研究含G/C的390 bp DNA片段與細胞覈提取物的結閤.結果:在中國北方人群α2A-AR基因中共髮現2處SNP.-1296位SNP等位基因G、C頻率分彆為0.61和0.39,基因型GG,GC,CC頻率分彆為0.34,0.54,0.13.EMSA結果錶明,細胞覈提取物與-1 296 bp為C的DNA片段可特異性結閤.結論:中國北方人群α2A-AR基因中存在2處SNP,調控區(-1 296位)SNP等位基因為C的DNA片段可與特異性蛋白質結閤,可能通過這一機製影響基因錶達.
목적:재중국인군신상선소α2A수체(α2A-AR)기인중수심단핵감산다태성(SNP)병연구기대기인표체적영향.방법:대α2A-AR기인진행서렬분석,수심SNP.종전혈중제취기인조DNA,확증포함G/C다태성위점적239 bp DNA편단.이용PCR-RFLP방법대중국북방인군α2A-AR기인-1296 bp위점SNP진행기인분형.이용응효조체방법연구함G/C적390 bp DNA편단여세포핵제취물적결합.결과:재중국북방인군α2A-AR기인중공발현2처SNP.-1296위SNP등위기인G、C빈솔분별위0.61화0.39,기인형GG,GC,CC빈솔분별위0.34,0.54,0.13.EMSA결과표명,세포핵제취물여-1 296 bp위C적DNA편단가특이성결합.결론:중국북방인군α2A-AR기인중존재2처SNP,조공구(-1 296위)SNP등위기인위C적DNA편단가여특이성단백질결합,가능통과저일궤제영향기인표체.
Objective: To scan single nucleotide polymorphism ( SNP ) in Chinese alpha-2A adrenergic receptor( α2A-AR )gene and study the effcts of the SNP on the gene expression .Methods: The complete sequence of α2A-AR gene was analyzed with autonated DNA sequencer to scan SNPs. Genomic DNA was extracted from whole blood and a 239 bp fragmnent containing the G/C polymorphism was amplified with PCR using a pair of specific primers.PCR-RFLP was used to perform the genotyping of the SNP at the site- 1 296 bp of the people in the North of China. Electrophoresis mobility shift assay( EMSA )was used to study the binding of the 390bp fragments(- 1 414-1 025 bp) with G or C at the site-1 296 bp and nuclear extracts. Results: In our study, two SNPs werefound in α2A-AR gene. Allele frequeneies of the SNP at the site- 1 296 bp were 0.61 and 0.39 for G and C, and the genotype frequencies were 0.34,0.54 and 0.13 for GG, GC and CC respectively from the people in the North of China.In the EMSA, a specific binding appeared in the complex of nuclear extracts and DNA with C at- 1 296 bp. Conclusion: Two SNPs exist in a2A-AR gene from the people in the North of China, and DNA fragment with allele C of the SNP at the site- 1 296 bp could bind with a specific protein, which could influence the gene expression.
