title: [Construction and sequence analysis of a normalized full-length cDNA library of Dendrobium officinale].
abstract: OBJECTIVE
  In order to obtain functional genes, a normalized stems cDNA library was constructed from medicinal plant Dendrobium officinale.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Yunnan Agricultural University, Advantages of Chinese Medicinal Materials Standardization Planting in Yunnan Province Engineering Research Center, Kunming 650201, China.
authors.forename: Min
authors.initials: M
authors.lastname: Jiang
authors.forename: Jiang
authors.initials: J
authors.lastname: Wang
authors.forename: Guo-Song
authors.initials: GS
authors.lastname: Wen
authors.forename: Shao-Zhong
authors.initials: SZ
authors.lastname: Xu
authors.forename: Ying-Hong
authors.initials: YH
authors.lastname: Zha
authors.forename: Tian-Ju
authors.initials: TJ
authors.lastname: Rong
authors.forename: Xiong
authors.initials: X
authors.lastname: Qian
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D031667:Dendrobium / Q000235:genetics*; D015723:Gene Library; D008969:Molecular Sequence Data; D010946:Plants, Medicinal / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger