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利用EcoRⅠ和HpaⅡ/MspⅠ双酶切建立适合于银杏基因组的甲基化敏感扩增多态性(methylation-sensitive amplification polymorphism,MSAP)分析体系,在全基因组水平检测银杏DNA甲基化修饰水平、模式及位点等表观遗传信息。结果显示,从54对MSAP选扩引物中,选出16对MSAP引物组合,共扩增产生454条清晰可辨且可重复的DNA条带,平均每对引物扩增获得28.38条带。在全部扩增位点中,检测到甲基化位点200个,CCGG/GGCC位点甲基化修饰比例为44%。部分银杏基因组甲基化修饰位点进行回收,最终分离了18条存在甲基化修饰的基因组DNA序列。BLASTn比对分析表明,银杏基因组中包括转录调控因子、反转录转座子、通道蛋白、启动子结合蛋白、蛋白激酶等在内的多种类型的DNA序列中均存在DNA甲基化修饰现象。
The methylation-sensitive amplification polymorphism (MSAP) analysis system suitable for Ginkgo biloba genome was established by EcoRⅠ and HpaⅡ / MspⅠ double digestion, and the level of DNA methylation was detected at the whole genome level. The pattern And sites such as epigenetic information. The results showed that of the 54 pairs of MSAP primers, 16 pairs of MSAP primers were selected and co-amplified 454 clear and repeatable DNA bands with an average of 28.38 bands per pair of primers. Among all the amplification sites, 200 methylation sites were detected, and the percentage of methylation in CCGG / GGCC site was 44%. Part of ginkgo genome methylation modified sites were recovered, and finally isolated 18 methylated modified genomic DNA sequences. BLASTn analysis showed that DNA methylation modification existed in many types of DNA sequences including transcriptional regulators, retrotransposons, channel proteins, promoter-binding proteins and protein kinases .