期刊文献+

小菜蛾PGRP-S2基因的克隆表达及功能分析 被引量:1

Cloning,expression and functional analysis of PGRP-S2 gene in Plutella xylostella
下载PDF
导出
摘要 肽聚糖识别蛋白(peptidoglycan recognition protein,PGRP)对于昆虫来说是一种高度保守的病原识别蛋白。为阐明PGRP-S2在小菜蛾Plutella xylostella抵抗病原微生物过程中的作用,本研究结合RT-PCR和RACE技术克隆得到小菜蛾PGRP-S2基因的cDNA全长序列,命名为PGRP-S2(GenBank登录号:MG570190)。生物信息学分析结果表明,PGRP-S2的开放阅读框为588 bp,编码195个氨基酸;蛋白质预测分子量为21.46 kDa,理论等电点为8.46;编码蛋白具有PGRP超家族保守结构域和酰胺酶结构域,是典型的肽聚糖识别蛋白,包含一条信号肽,不存在跨膜结构;同源序列比对和系统进化树分析表明PGRP-S2与家蚕Bombyx mori的BmPGRP-S 1进化距离最近。利用大肠杆菌Escherichia coli BL21(DE3)高效表达重组蛋白PxPGRP-S2,利用倒置显微镜及平板涂布观察重组蛋白对大肠杆菌E.coli和金黄色葡萄球菌Staphylococcus aureus的作用,结果表明PxPGRP-S2蛋白能够与两种细菌发生结合并凝集细菌,但不具备直接杀菌功能。本研究为进一步研究基于PGRP-S2介导的小菜蛾免疫防御反应提供基础。 Peptidoglycan recognition protein(PGRP)is a highly conserved pathogen recognition protein for insects.In order to elucidate the role of PGRP-S2 in the resistance of P.xylostlla to pathogenic microorganisms,the full-length cDNA of PGRP-S 2 gene of P.xylostlla was obtained by RT-PCR and RACE,and named as PGRP-S 2(GenBank accession:MG570190).According to bioinformatics analysis,with an open reading frame of 588 bp,PGRP-S 2 encodes 195 amino acids,the predicted molecular weight of the protein is about 21.46 kDa,and the theoretical isoelectric point is 8.46.The encoded protein has a conserved domain of PGRP superfamily and amidase domain,which are typical of peptidoglycan recognition proteins,and contains a signal peptide without transmembrane structure;homologous sequence alignment and phylogenetic tree analysis showed that PGRP-S 2 was closest to BmPGRP-S 1 of Bombyx mori.The recombinant protein PxPGRP-S2 was highly expressed in Escherichia coli BL21(DE3).The interaction between the recombinant protein PxPGRP-S2 with E.coli and Staphylococcus aureus was observed by an inverted microscope and plate coating.The results showed that PxPGRP-S2 protein could be bind to both bacteria and agglutinate bacteria,but not could kill them directly.The study would lay a foundation for the prevention and further study for the PGRP-S2-mediated immune defense responses in P.xylostella.
作者 杜少萱 吴程 苏月华 杨梅 DU Shao-Xuan;WU Cheng;SU Yue-Hua;YANG Mei(Fujian Normal University,School of Life Sciences,Fuzhou 350100,China)
出处 《环境昆虫学报》 CSCD 北大核心 2020年第2期410-418,共9页 Journal of Environmental Entomology
基金 福建省自然科学基金(2018J01726)。
关键词 小菜蛾 肽聚糖识别蛋白 原核表达 凝集试验 Plutella xylostella peptidoglycan recognition protein prokaryotic expression agglutination test
  • 相关文献

参考文献2

二级参考文献30

  • 1侯利霞,翟培,施用晖,乐国伟.不同细菌对家蝇幼虫抗菌蛋白/肽的诱导效应[J].昆虫知识,2006,43(6):827-831. 被引量:7
  • 2Steiner H. Peptidoglycan recognition proteins : on and off switches for innate immunity [ J ] . Immunol Rev, 2004, 198 : 83-96.
  • 3Dziarski R. Recognition of bacterial peptidoglycan by the innate immune system [ J ] . Cell Mol Life Sci, 2003, 60 : 1793-1804.
  • 4Ligoxygakis P, Pehe N, Hoffmann JA, et al. Activation of Drosophila toll during fungal infection by a blood serine protease [ J ] . Science, 2002, 297 : 114-116.
  • 5Michel T, Reichhart JM, Hoffmann JA, et al. Drosophila toll is activated by gram positive bacteria through a circulating peptidoglycan recognition protion [ J ] . Nature, 2001, 414 : 756- 759.
  • 6Chen KK, Liu C, He Y, et al. A short-type peptidoglycan recognition protein from the silkworm : expression, characterization and involvement in the prophenoloxidase activation pathway [ J ] . Dev Comp Immunol, 2014, 45 : 1-9.
  • 7Yang DQ, Su ZL, Qiao C, et al. Identification and characterizattion of two peptidoglycan recognition proteins with zinc-dependent antibacterial activity from the cotton Bollworm, Helicoverpa armigera [ J ] . Dev Comp Immunol, 2013, 39 : 343-351.
  • 8Iizuka M, Nagasaki T, Takahashi KG, et al. Involvement of pacific oyster CgPGRP-S1S in bacterial recognition, agglut!nation and granulocyte degranulation [ J ] . Dev Comp Immunol, 2014, 43 : 30-34.
  • 9Kurata S. Peptidoglycan recognition proteins in Drosophila Immu- nity [ J ]. Dev Comp Immunol, 2014, 42 : 36-41.
  • 10Lemaitre B, Hoffmann J. The host defense of Drosophila melanog- aster [ J ] . Annu Rev Immunol, 2007, 25 : 697-743.

共引文献7

同被引文献9

引证文献1

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部