Construction and sequence analysis of a normalized full-length cDNA library of Dendrobium officinale / 中国中药杂志
China Journal of Chinese Materia Medica
;
(24): 504-510, 2013.
Artigo
em Chinês
| WPRIM
| ID: wpr-318667
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>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plantas Medicinais
/
RNA Mensageiro
/
Dados de Sequência Molecular
/
Sequência de Bases
/
Biblioteca Gênica
/
Clonagem Molecular
/
Análise de Sequência de DNA
/
DNA Complementar
/
Dendrobium
/
Genética
Idioma:
Chinês
Revista:
China Journal of Chinese Materia Medica
Ano de publicação:
2013
Tipo de documento:
Artigo
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