Construction and sequence analysis of a normalized full-length cDNA library of Dendrobium officinale

• Published in: Zhongguo zhongyao zazhi Vol. 38; no. 4; p. 504
• Main Authors: Jiang, Min, Wang, Jiang, Wen, Guo-Song, Xu, Shao-Zhong, Zha, Ying-Hong, Rong, Tian-Ju, Qian, Xiong
• Format: Journal Article
• Language: Chinese
• Published: China 15.02.2013
• Subjects: Base Sequence; Cloning, Molecular; Dendrobium - genetics; DNA, Complementary - biosynthesis; DNA, Complementary - genetics; Gene Library; Molecular Sequence Data; Plants, Medicinal - genetics; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sequence Analysis, DNA - methods
• ISSN: 1001-5302
• DOI: 10.4268/cjcmm20130408

In order to obtain functional genes, a normalized stems cDNA library was constructed from medicinal plant Dendrobium officinale. 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. 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. 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.