Library construction of cDNA and SSR analysis of partial ESTs for stem and leaf of Betula luminifera.
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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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