新型花生内源特异参照基因的开发与应用研究  

Validation of a peanut-specific gene chi2.2 as an endogenous reference gene in qualitative PCR detection of peanut-derived products

作　　者：张丽[1,2] 吴刚[2] 武玉花[2] 肖玲[2] 曹应龙[2] 卢长明[2] 

机构地区：[1]中南民族大学生命科学院,湖北武汉430073 [2]中国农业科学院油料作物研究所,湖北武汉430062

出　　处：《中国油料作物学报》2008年第4期411-416,共6页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金(30700666)

摘　　要：一个物种的内源特异参照基因是该物种区别其它物种的特异性标志之一。本研究通过对GeneBank中花生基因信息的检索分析,筛选出花生几丁质酶基因chi2.2作为该物种内源特异参照基因的候选基因。通过对该基因的特定区域设计引物,建立了花生产品特异性定性PCR检测方法。利用该方法分别对16个花生品种的DNA进行PCR扩增,均能扩增出81bp的片段。利用相同引物分别对其他油料作物、粮食作物和蔬菜等(油菜,大豆,芝麻,向日葵,油棕,棉花,水稻,玉米,长豆角,豌豆,土豆,萝卜,辣椒,茄子,拟南芥,烟草)的DNA进行PCR扩增,均未发现特异性扩增产物。结果表明,本研究建立的花生产品检测方法具有很好的物种特异性,且其灵敏度达到0.05ng基因组DNA。该方法可用于花生源成分鉴定,如确定食品中是否含有花生源成分,本研究为识别掺假使杂和保障食品安全提供有效手段。A species-specific endogenous reference gene system was developed for polymerase chain reaction （PCR）-based analysis in peanut （Arachis hypogaea L. ） by targeting the chitinase gene chi2.2. The primers were elaborated for qualitative PCR assay. The amplified product was 81 base pairs in size. The specificity was assessed on 16 different peanut cultivars, other oil crops （ rapeseed, soybean, sesame, sunflower, oil palm and cotton）, vegetables （ carob, pea, potato, radish, pepper, eggplant） and other plant species （ maize, rice ,A. thaliana and tobacco）. Peanut-derived DNA was the only one amplified by the established PCR assay. The sensitivity of this method was assessed to be 0.05ng of peanut genomic DNA. This method proved to be highly specific and sensitive. It can be used to detect peanut-derived products, such as to determine the peanut components in edible oils, food and feedstuff, to identify the adulterations of peanut products and to use as quality control in transgenic peanut detection.

关 键 词：花生 内源特异参照基因 几丁质酶 PCR 食品安全 

分 类 号：S565.203[农业科学—作物学]