光皮桦茎叶cDNA文库构建及部分EST序列SSR分析
Library construction of cDNA and SSR analysis of partial ESTs for stem and leaf of Betula luminifera.
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摘要: 以光皮桦茎叶组织为材料，构建了cDNA文库。初级文库滴度为1.5×106pfu/mL，重组率达97.3%，插入片段大小在0.5～3.0 kb之间，平均长度约为1.3 kb，表明所构建的文库质量较高，可用于后续基因克隆及基因表达谱的研究。利用微卫星查找软件对获得的224条EST序列进行微卫星位点搜寻及其丰度、分布比较，发现47条序列含微卫星位点60个，占全部EST序列的26.80%；在所有SSRs中二碱基重复最多，为42个，占总数的70.00%，含三、四碱基重复分别占总数的28.30%和1.70%。通过对光皮桦EST序列中微卫星位点信息的发掘分析，为有针对性地设计ESTSSR引物、进行遗传多样性分析奠定了基础。Abstract: A cDNA library of stem and leaf from Betula luminifera was constructed. The primary titer of cDNA library was about 1.5×106 pfu/mL, its recombinant efficiency reached 97.3%, and the size of insert DNA fragments ranged from 0.5 to 3.0 kb, with an average of 1.3 kb. The results indicated that it was a higherquality cDNA library, and could be used in gene cloning and gene expression profile analysis. Distribution and frequency of SSRs were analyzed in 224 nonredundant ESTs from B. luminifera cDNA library, using online searching software. The results showed that 60 SSRs distributed in 47 EST sequences, accounting for 26.80% of all ESTs. Dinucleotide would be the major repeat types, accounting for 70.00% of the total number of acquired SSRs. The trinucleotide and tetranucleotide repeats accounted for 28.30% and 1.70% respectively. This research might lay the foundation for designing the targeted ESTSSR primers and genetic diversity analysis by mining the information of EST-SSR loci in B. luminifera EST sequence data.
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Keywords:
- Betula luminifera /
- cDNA library /
- SSR analysis
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