Construction and sequence analysis of a normalized full-length cDNA library of Dendrobium officinale / 中国中药杂志

ABSTRACT

<p><b>OBJECTIVE</b>In order to obtain functional genes, a normalized stems cDNA library was constructed from medicinal plant Dendrobium officinale.</p><p><b>METHOD</b>SMART (switching mechanism at 5' end of RNA transcript) cDNA synthesis combined with DSN (duplex-specific nuclease) normalization was applied to construct the normalized full-length cDNA library of D. officinale.</p><p><b>RESULT</b>The titer of cDNA library was about 1.3 x 10(6) cfu x mL(-1) and the average insertion size was about 1.5 kb with high recombination rate (93.9%). Random selected 163 positive clones were sequenced at single side. Bio-information analysis indicated that 147 from 150 high-quality unique sequences matched corresponding homologous proteins, and they participated in various biological processes based on GO (gene ontology). There were 8 clones with complete coding sequence, which presumed to be full-length genes.</p><p><b>CONCLUSION</b>These results showed preliminarily that we successfully constructed a normalized full-length cDNA library of D. officinale which could be used to screen the functional genes related to metabolic pathways of medicinal ingredients.</p>