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Volume 24 Issue 3
May  2003
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Article Navigation >  Zoological Research  > 2003  >  24(3): 215-219
SHEN Ben-chang, JIN Zhi-ping, ZHAO De-xiu, QIAO Chuan-ling. Construction of a Full-length cDNA Library of the Diamondback Moth,Plutella xylostella. Zoological Research, 2003, 24(3): 215-219.
Citation: SHEN Ben-chang, JIN Zhi-ping, ZHAO De-xiu, QIAO Chuan-ling. Construction of a Full-length cDNA Library of the Diamondback Moth,Plutella xylostella. Zoological Research, 2003, 24(3): 215-219.
shu

Construction of a Full-length cDNA Library of the Diamondback Moth,Plutella xylostella

  • 1.

    State Key Laboratory of Integrated Management of Pest Insects and Rodents,Institute of Zoology,the Chinese Academy of Sciences,Beijing 100080,China;2.Institute of Botany,the Chinese Academy of Sciences,Beijing 100093,China

  • Received Date: 1900-01-01
  • Rev Recd Date: 1900-01-01
  • Publish Date: 2003-06-22
    Abstract
  • In order to understand the molecular mechanism of the pesticide resistance of diamondback moth,Plutella xylostella,we used the SMART (switching mechanism at 5 end of RNA transcript) technology to construct full-length cDNA libraries.The PowerScript reverse transcriptase was used to synthesize and anchor the first-strand cDNA.Following reverse transcription,LD-PCR was performed using a modified oligo (dT) and an anchor primer to enrich the cDNA population for full-length sequences.The unamplified cDNA library contained 2.6×10[6] independent clones,in which the recombinant clones with an average insert size of 1.6 kb was > 96%.The titer of the amplified library was 2.4×10[10] pfu/mL.PCR analysis of 60 randomly selected clones showed that 65% of cDNA clones contained inserts of above 1.0 kb.From the cDNA library,a full-length ribosomal protein L27A cDNA with 5 and 3 untranslated regions was isolated.
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