Construction and sequence analysis of a normalized full-length cDNA library of Dendrobium officinale.
- Author:
Min JIANG
1
;
Jiang WANG
;
Guo-Song WEN
;
Shao-Zhong XU
;
Ying-Hong ZHA
;
Tian-Ju RONG
;
Xiong QIAN
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Cloning, Molecular; DNA, Complementary; biosynthesis; genetics; Dendrobium; genetics; Gene Library; Molecular Sequence Data; Plants, Medicinal; genetics; RNA, Messenger; genetics; metabolism; Sequence Analysis, DNA; methods
- From: China Journal of Chinese Materia Medica 2013;38(4):504-510
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEIn order to obtain functional genes, a normalized stems cDNA library was constructed from medicinal plant Dendrobium officinale.
METHODSMART (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.
RESULTThe 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.
CONCLUSIONThese 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.
