磁珠富集法开发毛竹SSR标记引物

Development of Polymorphic Simple Sequence Repeats Markers in Phyllostachys edulis by Magnesphere

摘要: 用磁珠富集法筛选毛竹基因组DNA的SSR分子标记,利用磁珠法对毛竹基因组DNA的AFLP片段进行了富集,分别构建了富含GT、AG、CCA重复基元的富集文库,利用克隆PCR法对文库进行筛选,检测菌落数分别为1080、620、630个,阳性菌落中含目的重复的序列分别为137、73、41个,富集效率分别为12.7%、11.8%、6.5%。双碱基重复的富集效率高于三碱基富集效率。根据获得的序列结果,设计了53对SSR引物,其中31对引物在毛竹中成功扩增出目的大小的条带,扩增成功率为58.9%。

Abstract: The objective of this work is to seek SSR markers for Phyllostachys edulis. Magnesphere method was used to conentrate SSR containing sequences from Ph. edulis Genomic DNA AFLP fragments. Three SSR-enriched libraries (GT, AG, and CCA) were constructed. The Clone-PCR method was used to screen positive clones, 1 080, 620, 630 clones were screened in GT, AG, CCA libraries, and 137, 73, 41 SSR-containing sequences were obtained, at concentration rate of 12.7%, 11.8%, 6.5% respectively. The result showed that the concentration rate of dinucleotide repeat libraries is higher than trinucleotide libraries. After sequences analyzing, 53 pairs of SSR primers were designed, 31 of which amplified the objective fragment in Ph. edulis at the rate of 58.9%.