Molecular cloning and characterization of the lipopolysaccharide and beta-1,3-glucan binding protein from red claw crayfish, Cherax quadricarinatus

文献类型： 外文期刊

第一作者： Hou, Libo

作者： Hou, Libo;Liu, Xiaoqian;Jing, Yunting;Zhang, Jia;Gu, Wei;Meng, Qingguo;Wang, Wen;Hou, Libo;Liu, Xiaoqian;Jing, Yunting;Zhang, Jia;Gu, Wei;Meng, Qingguo;Wang, Wen;Ding, Zhengfeng;Huang, Yanqing

作者机构：

关键词： Lipopolysaccharide and beta-1;3-glucan binding protein (LGBP);Molecular cloning;Immune response;Cherax quadricarinatus

期刊名称：AQUACULTURE （ 影响因子：4.242； 五年影响因子：4.723 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： A pattern recognition protein (PRP), lipopolysaccharide and beta-1,3-glucan binding protein (LGBP), cDNA was cloned from the hemocytes of red claw crayfish, Cherax quadricarinatus by the techniques of homology cloning and RACE. Analysis of nucleotide sequence revealed that the full-length cDNA of 1456 bp has an open reading frame of 1095 bp encoding a protein of 364 amino acids including a 15 amino acid signal peptide. The calculated molecular mass of the mature protein (349 aa) was 39.92 kDa with an estimated pI of 4.50. Sequence comparison of the deduced amino acid sequence of C. quadricarinatus LGBP (named as CqLGBP) showed a high identity of 82%, 77%, 76% and 74% with Procambarus clarkii LGBP, Homarus gammanus BGBP, Penaeus monodon BGBP and Litopenaeus stylirostlis LGBP, respectively. The CqLGBP sequence contains: (1) two putative integrin-binding motifs, (2) a glucanase motif, (3) two putative N-glycosylation sites, (4) one protein kinase C phosphorylation site, and (5) a putative recognition motif for beta-1,3-linkage of polysaccharides. The recombinant CqLGBP were expressed in Escherichia coli BL21 with pGEX-6P-2 expression vector. The titer of rabbit anti-CqLGBP serum was above 1:12,800. Microorganism binding assay showed CqLGBP binds two kinds of crustacean pathogens Spiroplasma eriocheiris and Aeromonas hydrophila. Quantitative RT-PCR method was used to quantify the variation of mRNA transcription level during artificial infection with S. eriocheiris and A. hydrophila. A significant enhancement of CqLGBP transcriptions was apparent post-injection in response to bacterial infection compared to the control group. These data should be helpful to better understand the function of CqLGBP in the crayfish immune system. (C) 2015 Elsevier B.V. All rights reserved.

分类号： S9

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