华北农学报 ›› 2008, Vol. 23 ›› Issue (6): 72-76. doi: 10.7668/hbnxb.2008.06.016

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芜菁夜蛾斯氏线虫G26侵染期幼虫cDNA文库的构建及表达序列标签分析

马娟1, 张军鸽1,2, 李秀花1, 李敏权2, 陈书龙1   

  1. 1. 河北省农林科学院, 植物保护研究所, 河北省农业有害生物综合防治工程技术研究中心, 河北, 保定, 071000;
    2. 甘肃农业大学, 甘肃, 兰州, 730070
  • 收稿日期:2008-07-10 出版日期:2008-12-28
  • 作者简介:马娟(1981-),女,河北保定人,研究实习员,硕士,主要从事农业线虫研究。
  • 基金资助:
    国家自然科学基金(30671409);公益性行业科技专项(nyhyzx07-050-12);国家科技支撑计划(2006BAI06A10-3)

Construction of the cDNA Library from IJs of Steinernema feltiae G26 and Analysis of Expressed Sequence Tags

MA Juan1, ZHANG Jun-ge1,2, LI Xiu-hua1, LI Min-quan2, CHEN Shu-long1   

  1. 1. Institute of Plant Protection, Hebei Academy of Agriculture and Forestry Sciences IPM, Centre of Hebei Province, Baoding 071000, China;
    2. Gansu Agricultural University, Lanzhou 730070, China
  • Received:2008-07-10 Published:2008-12-28

摘要: 本研究以芜菁夜蛾斯氏线虫Steinernema feltiae G26的侵染期幼虫为材料,利用SMART (Switching mechanism at ’end of RNA transcript)技术,构建了其cDNA文库,得到原始文库的库容为3.7×108 cfu/mL。随机挑取40个克隆,测序获得386条质量较好的序列,插入片段多在0.3~3 kb之间。将EST序列进行同源性比对, EST中有注释信息的占9.92%,无注释信息的4.08%。蛋白功能分析表明所得EST包括核糖体蛋白类,调节因子,信号传导相关基因,代谢酶类等。此全长cDNA文库的构建成功,为阐明侵染期抗逆性幼虫形成与发育机制以及抗逆代谢机理等提供了重要理论依据。

关键词: 昆虫病原线虫, SMART技术, cDNA文库

Abstract: Entomopathogenic nematodes(EPN)is a new type of bio-pesticide which has great potential for controlling many important agricultural pests.The infective juveniles(IJ s)play an important role in its commercialization.A full length cDNA library from IJ s of Steinernema feltiae G26 was constructed using SMART techniques.The titer of primary cDNA library was 3.7×108cfu/mL.Sequencing and sequence similarity searches were performed to analyze theprofile of expressed sequence tags(ESTs).386 qualified sequences were obtained from 450 clones,the insert size ranged from 0.3-3.0 kb.ESTs sequences similarity was compared in non-redundancies database of GenBank.The results showed that 4.08 % ESTs had none homology and these which had high homology included ribosomal protein,translation elongation factor,signal transduction gene,metabolize enzyme and so on.Therefore,a cDNA library ofSteinernema feltiaeG26 has been successfully constructed.It provides basic information for understanding the mechanisms on formation,development and persistence of IJ s.

Key words: Entomopathogenic nematodes, SMART techniques, cDNA library

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引用本文

马娟, 张军鸽, 李秀花, 李敏权, 陈书龙. 芜菁夜蛾斯氏线虫G26侵染期幼虫cDNA文库的构建及表达序列标签分析[J]. 华北农学报, 2008, 23(6): 72-76. doi: 10.7668/hbnxb.2008.06.016.

MA Juan, ZHANG Jun-ge, LI Xiu-hua, LI Min-quan, CHEN Shu-long. Construction of the cDNA Library from IJs of Steinernema feltiae G26 and Analysis of Expressed Sequence Tags[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(6): 72-76. doi: 10.7668/hbnxb.2008.06.016.