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对来源于NCBI公共数据库的非冗余3087条毛竹(Phyllostachys edulis)EST序进行简单重复序列SSR搜索发现:在401条毛竹EST序列中共查找到408个SSR位点,平均6.8kb EST序列中含有1个SSR。其中二核苷酸和三核苷酸重复是毛竹EST-SSR的主要重复类型,分别占SSR总数的43.14%和49.75%。SSR的不同重复基序中,最常见的重复基序是:(AG)n,占37.01%,其余出现频率较高的依次为(TC)n、(AGC)n、(GAG)n和(CGC)n。根据搜索结果设计了182条毛竹EST-SSR引物,以12个不同种属的竹子DNA为模板,对引物进行了稳定性、通用性的验证与评价。结果表明有42对引物有较清晰且稳定的目标扩增产物,其中39对引物的产物存在多态性,说明设计开发的毛竹的EST-SSR标记是可行且有效的。
A simple repeat SSR search of non-redundant 3087 Phyllostachys edulis EST sequences originated from NCBI public database found that 408 SSR sites were found among 401 Phyllostachys pubescens EST sequences, and the average 6.8 kb EST sequence contained 1 SSR. The dinucleotide and trinucleotide repeats were the major repeat types of EST-SSR, which accounted for 43.14% and 49.75% of the total number of SSR respectively. Among the different repeat motifs of SSR, the most common repeat motif is (AG) n, accounting for 37.01%. The other most frequently occurring recurring motifs are (TC) n, (AGC) n, (GAG) n and n. Based on the search results, 182 EST-SSR primers were designed and used to validate and evaluate the stability and versatility of the primers from 12 different species of bamboo DNA. The results showed that 42 pairs of primers had a clearer and more stable target amplification products, of which 39 pairs of primers had polymorphism, which indicated that the EST-SSR marker of the developed bamboo was feasible and effective.