人参植物高质量RNA的分离及其皂苷生物合成相关新基因GBR6的表达分析

Isolation of High-quality RNA and Expression Analysis of GBR6 Gene Involved in Ginsenoside Biosynthesis from Panax ginseng Plant Tissues

为从富含多糖的人参植物组织中分离出高质量的RNA,使用改进的两种异硫氰酸胍法,可从人参植物根组织中提取到较高产量和质量的总RNA。每克新鲜组织的RNA产量在80～110μg之间;经琼脂糖凝胶电泳分析,可见到28S和18SrRNA两条完整、清晰的条带;紫外分光光度法测得的A_(260)/A_(280)比值位于1.8～2.0。而且,使用该改良法分离的RNA,可广泛用于检测基因表达的RT-PCR与Northern印迹杂交分析以及cDNA文库构建等植物分子生物学研究。

To find a simple and efficient method for isolating total RNA from Panax ginseng plant tissues rich in polysaccharides, two modified acid guanidinium thiocynate extraction methods were tested with fresh Panax ginseng roots tissues. These techniques, compared to other methods. gave better RNA yield in the range of 80-110 μg per gram fresh weight of tissue and higher quality with A260/A 280 ratios in the range of 1. 8 and 2. 0 and distinct 28S and 18S rRNA bands. Further, the total RNA obtained from the roots of Panax ginseng were suitable for a subtractive cDNA library construction, RT-PCR and Northern blotting hybridization assays. .