Molecular cloning and characterisation of a pattern recognition molecule, lipopolysaccharide- and β-1,3-glucan binding protein (LGBP) from the white shrimp Litopenaeus vannamei
Molecular cloning and characterisation of a pattern recognition molecule, lipopolysaccharide- and β-1,3-glucan binding protein (LGBP) from the white shrimp Litopenaeus vannamei
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DOI：
10.1016/j.fsi.2004.08.002
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发表时间：
2005-04-01
影响因子：
4.7
通讯作者：
Chen, JC
中科院分区：
文献类型：
--
作者：
Cheng, WT;Liu, CH;Chen, JC
利用寡核苷酸引物和RT-PCR技术，从白色凡纳滨对虾血细胞和肝胰腺中克隆了脂多糖和β-1，3-葡聚糖结合蛋白（LGBP）cDNA。用cDNA末端快速扩增RACE法分离3 ′-和5 ′-区。核苷酸序列分析表明，该cDNA克隆具有一个1101 bp的开放阅读框架，编码367个氨基酸的蛋白质，包括一个17个氨基酸的信号肽。成熟蛋白（350个氨基酸）的计算分子量为39.92 kDa，估计pI为4.37。在LGBP中观察到两个推定的整合素结合基序（细胞粘附位点），RGD（Arg-Gly-Asp）和多糖的β-（1 -> 3）键的潜在识别基序。序列比较表明，LGBP推导的氨基酸序列与L.与南美白对虾（Litopenaeus stylirostris LGBP）、斑节对虾（Penaeus monodon BGBP）和小龙虾（Pacifstacus leniusculus LGBP）的相似性分别为95%、92%和61%。实时荧光定量RT-PCR分析结果表明，LGBP在血细胞中的转录水平与对照组相比有显著性差异。在注射溶藻弧菌后3和6 h，凡纳滨对虾增加。(C)2004爱思唯尔有限公司保留所有权利。
A lipopolysaccharide- and beta-1,3-glucan binding protein (LGBP) cDNA was cloned from the haemocyte and hepatopancreas of white shrimp Litopenaeus vannamei using oligonucleotide primers and RT-PCR. Both 3'- and 5'-regions were isolated by rapid amplification of cDNA end RACE method. Analysis of nucleotide sequence revealed that the cDNA clone has an open reading frame of 1101 bp encoding a protein of 367 amino acids including a 17 amino acid signal peptide. The calculated molecular mass of the mature proteins (350 amino acids) is 39.92 kDa with an estimated pI of 4.37. Two putative integrin binding motifs (cell adhesion site), RGD (Arg-Gly-Asp) and a potential recognition motif for beta- (1 --> 3) linkage of polysaccharides were observed in the LGBP. Sequence comparison showed that LGBP deduced amino acid of L. vannamei has an overall similarity of 95%, 92% and 61 % to that of blue shrimp Litopenaeus stylirostris LGBP, tiger shrimp Penaeus monodon BGBP and crayfish Pacifstacus leniusculus LGBP, respectively. Quantitative real-time RT-PCR analysis showed that LGBP transcript in haemocyte of L. vannamei increased in 3- and 6-h post Vibrio alginolyticus injection. (C) 2004 Elsevier Ltd. All rights reserved.