Variation in dinucleotide (GT) repeat sequence in the first exon of the STAT6 gene is associated with atopic asthma and differentially regulates the promoter activity in vitro
Variation in dinucleotide (GT) repeat sequence in the first exon of the STAT6 gene is associated with atopic asthma and differentially regulates the promoter activity in vitro
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DOI：
10.1136/jmg.2003.015842
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发表时间：
2004-07-01
影响因子：
4
通讯作者：
Huang, SK
中科院分区：
文献类型：
--
作者：
Gao, PS;Heller, NM;Huang, SK
材料和方法研究人群和基因分型15名随机选择的白色美国人和214名白色英国人进行了研究。英国主题选择的细节已经在前面描述过了。6所有哮喘受试者均经医生诊断为哮喘，伴有（a）需要持续治疗的反复呼吸困难和胸闷，（B）有喘息记录，（c）有不稳定气流阻塞，连续呼气峰流速变异性为。百分之三十特应性定义为皮肤点刺试验阳性。5 mm和特异性IgE（。3.5 kU/l）对抗空气过敏原屋尘螨或草花粉。通过CAP ELISA系统（Pharmacia，Uppsala，Sweden）进行总血清IgE的测量。使用商业试剂盒（IsoQuick，Microbe Corp，加登格罗夫，美国）提取DNA样品，并对STAT 6基因的碱基对272至+115（相对于转录起始位点; GenBank AF 067575）进行PCR扩增。使用引物对59-TCTTCTCTGTGTCGTCAGAG和59-CAACACCACCACCACCCCGA来扩增该片段，然后通过基于激光的377自动测序仪和GeneScan软件（Applied Biosystem）进行基因分型。
MATERIALS AND METHODS Study population and genotyping Fifteen randomly selected white American and 214 white British subjects were studied. Details of the British subject selection have been previously described. 6 All the asthmatic subjects had physician diagnosed asthma with (a) recurrent breathlessness and chest tightness requiring ongoing treatment,(b) documented wheeze, and (c) documented labile airflow obstruction with variability in serial peak expiratory flow rates of. 30%. Atopy was defined as a positive skin prick test of. 5 mm and specific IgE (. 3.5 kU/l) against either of the aeroallergens house dust mite or grass pollen. Measurement of total serum IgE was carried out by the CAP ELISA system (Pharmacia, Uppsala, Sweden). DNA samples were extracted using a commercial kit (IsoQuick, Microprobe Corp, Garden Grove, USA), and subjected to PCR amplification of base pairs 272 to+ 115 (relative to the transcription start site; GenBank AF067575) of the STAT6 gene. Primers pairs 59-TCTTCTCTGTGTCGTCAGAG and 59-CAACACCACCACCACCCCGA were used to amplify the fragment, which was then genotyped by a laser based 377 automated sequencer and GeneScan software (Applied Biosystems).