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褐飞虱中肠内膜结合短肽P2S的筛选
  • ISSN号:0454-6296
  • 期刊名称:昆虫学报
  • 时间:2013.3.3
  • 页码:219-227
  • 分类:Q966[生物学—昆虫学]
  • 作者机构:[1]福建农林大学生物农药与化学生物学教育部重点实验室,福州350002
  • 相关基金:国家“863”计划项目(2011AAl0A203);国家自然科学基金项目(31071745);高等学校博士学科点专项科研基金(20093515110010)
  • 相关项目:苏云金杆菌Cry毒素抗假眼小绿叶蝉的定向筛选与改造
中文摘要:

【目的】筛选与褐飞虱Nilaparvata lugens中肠内膜相结合的短肽,探索能阻断褐飞虱传播水稻病毒的新方法。【方法】通过改良的膜饲喂法和噬菌体短肽文库展示技术,模拟褐飞虱正常取食的行为,筛选能够与其中肠内膜结合的短肽。将筛选到的短肽与加强绿色荧光蛋白进行融合表达,经纯化后,将融合蛋白饲喂给褐飞虱若虫,通过荧光显微镜观察饲喂后若虫的中肠。【结果】使用改良的膜饲喂法,16h后褐飞虱若虫的取食率达到91%,24h后达到95%。经形态学验证,确定了一种短肽与褐飞虱中肠内膜具有结合活性,并将其命名为P2S。【结论】改良了针对褐飞虱的膜饲喂法,筛选并验证了能够与褐飞虱中肠内膜结合的活性短肽P2S。研究结果为阻断相关的水稻病毒与褐飞虱中肠内膜间的相互作用研究提供了科学依据,也为褐飞虱新型膜穿孔毒素蛋白的研究提供了理论基础。

英文摘要:

[ Aim] To screen peptides which may bind to the midgut inner membrane in the rice brown planthopper, Nilaparvata lugens, so as to explore the new methods for blocking N. lugens to transmit rice viruses. [ Methods ] Trophic behavior of nymphs of the rice brown planthopper was imitated with improved membrane was fed to the fusion feeding method. Through the use of this feeding method, the phage peptide library solution planthopper nymphs to screen peptides which may bind to the midgut inner membrane. Then, expression of peptide and enhanced green fluorescent protein was conducted, and the purified fusion protein was fed to nymphs of the planthopper. To confirm the binding activity of screened peptide, the green fluorescence of fusion protein in the midgut of the nymphs was observed under the fluorescence microscope. [ Results] The feeding rates of nymphs of the planthopper detected by improved membrane feeding method at 16 h and 24 h after treatment reached 91% and 95%, respectively. By observing the green fluorescence from planthopper gut, one peptide, named P2S, was determined as the planthopper gut inner membrane binding peptide. [ Conclusion ] Membrane feeding method was well improved for the rice brown planthopper. A peptide P2S which may bind to the midgut inner membrane of the planthopper was effectively screened and its binding activity was confirmed. The results will provide a possibility of blocking interaction between rice viruses and the midgut inner membrane of the rice brown planthopper. Furthermore, the peptide P2S may be used for further study of the membrane insertion toxins binding to proper receptors in the midgut of the rice brown planthopper.

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期刊信息
  • 《昆虫学报》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学院
  • 主办单位:中国昆虫学会 中国科学院动物研究所
  • 主编:黄大卫
  • 地址:北京东中关村中国科学院动物研究所
  • 邮编:100101
  • 邮箱:kcxb@ioz.ac.cn
  • 电话:010-64807173 010-64807099
  • 国际标准刊号:ISSN:0454-6296
  • 国内统一刊号:ISSN:11-1832/Q
  • 邮发代号:2-153
  • 获奖情况:
  • 国家期刊方阵“双百期刊”
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),英国农业与生物科学研究中心文摘,美国剑桥科学文摘,美国生物科学数据库,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:16190