| 草地螟普通气味结合蛋白Ⅰ（Lsti-GOBP1）蛋白表达纯化及结合特性分析 |
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| 引用本文： | 孙红岩,尹姣,冯红林,李克斌,席景会,曹雅忠.草地螟普通气味结合蛋白Ⅰ（Lsti-GOBP1）蛋白表达纯化及结合特性分析[J].昆虫学报,2011,54(4):381-389. |
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| 作者姓名： | 孙红岩 尹姣 冯红林 李克斌 席景会 曹雅忠 |
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| 作者单位： | 吉林大学植物科学学院;中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室; |
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| 基金项目： | 公益性行业(农业)科研专项(201003079) |
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| 摘 要： | 气味结合蛋白在昆虫对寄主挥发性气味识别过程中有着重要的生理功能。本研究通过对草地螟Loxostege sticticalis L. 普通气味结合蛋白Ⅰ（Lsti-GOBP1）进行克隆及原核表达的基础上, 分离纯化得到体外重组的Lsti-GOBP1蛋白。并利用N-苯基-1-萘胺(N-phenyl-1-naphthylamine, 1-NPN）作为荧光探针研究了Lsti-GOBP1蛋白与醇类、 醛类、 酯、 烯等50种气味标样的结合特性, 结果表明Lsti-GOBP1可与其中35种气味化合物结合, 但只有1-己醇、 1-庚醇、 肉桂醛和莰烯4种气味标样能在20 μmol/L浓度（探针与蛋白结合的饱和浓度值）下将1-NPN从Lsti-GOBP1中替换50%, 其结合常数分别为8.997, 7.283, 7.289 和9.814 μmol/L。据此可以得出, Lsti-GOBP1蛋白的气味物质结合谱很广, 同时具有较强的特异性, 其中1-己醇、 1-庚醇、 肉桂醛、 莰烯等化合物在草地螟识别寄主植物气味物质的过程中起重要作用。
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| 关 键 词： | 草地螟 普通气味结合蛋白Ⅰ 蛋白表达 纯化 N-苯基-1-萘胺 荧光结合实验 |
| 收稿时间： | 2010-09-16; |
Expression, purification and binding characteristics of general odorant binding protein Ⅰ(GOBP1) from the meadow moth, Loxostege sticticalia (Linnaeus) (Lepidoptera: Pyralididae) |
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| SUN Hong-Yan,YIN Jiao,FENG Hong-Lin,LI Ke-Bin,XI Jing-Hui,CAO Ya-Zhong.Expression, purification and binding characteristics of general odorant binding protein Ⅰ(GOBP1) from the meadow moth, Loxostege sticticalia (Linnaeus) (Lepidoptera: Pyralididae)[J].Acta Entomologica Sinica,2011,54(4):381-389. |
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| Authors: | SUN Hong-Yan YIN Jiao FENG Hong-Lin LI Ke-Bin XI Jing-Hui CAO Ya-Zhong |
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| Institution: | SUN Hong-Yan1,2,YIN Jiao2,FENG Hong-Lin1,LI Ke-Bin2,XI Jing-Hui1,CAO Ya-Zhong2(1. College of Plant Science,Jilin University,Changchun 130062,China,2. State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China) |
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| Abstract: | Odorant-binding proteins (OBPs) are responsible for physiological function via perception of volatile odors. Here we reported the successful expression and purification of a general odorant binding protein Lsti-GOBP1 from Loxostege sticticalis L. Lsti-GOBP1 was expressed using Escherichia coli Rosetta (DE3) prokaryotic expression system,and then purified using Histrap HP column,and the protein functions of Lsti-GOBP1 were tested by fluorescent probe N-phenyl-1-naphthylamine (1-NPN) binding assays and compet... |
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| Keywords: | Loxostege sticticalis" target="_blank">Loxostege sticticalis')" href="#">Loxostege sticticalis general odorant binding protein Ⅰ(GOBP1) protein expression purification N-phenyl-1-naphthylamine fluorescent binding assay |
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