杜松烯合成酶作为转基因棉花PCR检测的内参照基因

Validation of a cotton specific gene,(+)-d-cadinene synthase,used as the endogenous reference gene for real-time PCR transgenic detection

根据棉属植物中棉酚合成的关键酶之一杜松烯合成酶[(+)dcadinenesynthase]基因序列,设计合成了该酶的实时荧光PCR的引物和TaqMan探针,经研究发现,此套引物探针能特异性检测海岛棉和陆地棉,有较高的检测灵敏度。同马铃薯、大椒、茄子、烟草、番茄、玉米、大豆、小麦、水稻和其它转基因作物无非特异性反应,可以作为转基因棉花检测的内参照。

Many countries have issued regulations to label the genetically modified organisms （GMOs）and their derived products.in this article, we reported the development of specific primers and probe for the cotton（Gossypium hirsutum） （ ＋ ）-d-cadinene synthase gene and PCR cycling conditions suitalbe for which use as an endogenous reference gene in PCR assays. Expect results were obtained when method was assayed with 7 different cotton varieties, and no amplification were observed when other species, such as wheat, maizes, tobecco, soybean, rice, tomato, pepper, eggplant were used as templates, which demoustmted the（ ＋ ）-d-cadinene synthase gene was cotton specific, had low heterogeneity among the tested cultivars. Therefore, this gene could be used as an endogenous reference gene of cotton in practial real time transgenic detection.