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>
Subject(s)
Base Sequence , Cloning, Molecular , DNA, Complementary , Genetics , Dendrobium , Genetics , Gene Library , Molecular Sequence Data , Plants, Medicinal , Genetics , RNA, Messenger , Genetics , Metabolism , Sequence Analysis, DNA , MethodsABSTRACT
<p><b>OBJECTIVE</b>To investigate the genetic relationships of Erigeron breviscapus at the molecular biology level.</p><p><b>METHOD</b>Thirty seven germplasm resources of E. breviscapus which collected from Yunnan, Sichuang and Guizhou province in 2005 were analyzed by Random amplified polymorphic DNA (RAPD) and cluster analysis based on NTSYS2.</p><p><b>RESULT</b>A total of 10 primers were screened, and 107 bands were amplified, among which 94 (87.85%) bands were found to be polymorphic. Thirty seven germplasm resources of E. breviscapus were clustered into 3 groups at genetic distance 0.36, the I group include in 9 germplasm resources collected from Mile, Qiubei, Luxi, Gejiu, and Yanshan of south east of Yunnan province; the II group included 8 germplasm resources collected from Gucheng, and shangrila of north west of Yunnan province, and Mile and Qiubei of south east of Yunnan province; the III group included in 20 germplasm resources collected from the center of Yunnan province, north east of Yunnan province, Sichuan province, and Guizhou province.</p><p><b>CONCLUSION</b>There were abundant genetic diversity in the germplasm resources of E. breviscapus, and the genetic relationships are closely related to geographical distance where they were collected.</p>