基于已获得的球囊菌转录组数据预测微卫星标记(SSR),并进行SSR位点的信息分析和SSR引物的挖掘.利用软件MISA对球囊菌转录组中42610条unigenes进行搜索,共预测出7968个SSRs,分布于5233条unigenes中,其中最主要的重复类型为三核苷酸重复(53.15%),其次为二核苷酸重复(32.32%)和四核苷酸重复(8.46%).二核苷酸重复中的基序主要是AG/CT(占总量的15.8%).针对所有的SSR位点,利用Primer Premier 5软件设计出6956对SSR特异性引物,随机选取20对引物对两个不同来源的球囊菌样品进行SSR位点扩增,共有6对引物成功扩增出符合预期的目的片段.研究结果表明,利用球囊菌转录组数据开发SSR引物是可行的.%To illustrate the molecular genetics and functional genomics of Ascosphaera apis, simple sequence markers ( SSRs) were predicted based on previous transcriptome data of A.apis, and SSR loci were analyzed and the specific SSR primers were developed. Firstly, 42610 unigenes assembled from RNA-seq data were interpreted by MISA software, and 7968 SSR loci were detected, they were distributed in 5233 unigenes. Among these SSR loci, tri-nucleotide repeats were the dominant repeat type, accounting for 53.15% of the total, which was followed by di-nucleotide repeats (32.32%) and tetra-nucleotide repeats (8.46%). The AG/CT motif was the majority (15.8%) in di-nucleotide repeats. In total, 6956 pairs of primers were designed from all SSRs. Among them, 20 SSR primer pairs were randomly selected and used to amplify the SSR loci in A.apis samples from 2 regions in China. It turned out that 6 pairs of primers were able to amplify the target fragments with expected sizes, indicating that large-scale development of A.apis SSR primers on the basis of transcriptome data is feasible.
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