三角帆蚌短型肽聚糖识别蛋白S3基因克隆及表达分析

发布时间：2019-04-07 18:51

【摘要】：肽聚糖识别蛋白(PGRPs)是机体先天免疫系统中的重要模式识别受体,研究对三角帆蚌的一种短型肽聚糖识别蛋白进行了克隆与表达分析。研究采用5′、3′RACE技术,对高通量转录组测序所得三角帆蚌PGRPS3基因(hc PGRPS3)片段分别进行了5′,3′末端克隆,经拼接得到hc PGRPS3 c DNA全长序列。采用多种分子生物学软件对hc PGRPS3 c DNA全长序列进行了特征分析,实时荧光定量(q RT-PCR)检测了其组织分布,及经肽聚糖(Peptidoglycan,PGN)和脂多糖(Lipopolysaccharide,LPS)刺激后其在肝胰腺中的基因表达变化。hc PGRPS3 c DNA全长1438 bp,开放阅读框为858 bp,编码285个氨基酸,预测分子量大小为32.3 ku,p H 7.0时的理论等点为7.98。序列中不存在信号肽与跨膜结构。氨基酸序列保守性分析表明,其他物种短型PGRP中,以夏威夷短尾鱿PGRP4与hc PGRPS3同源性最高(51%)。q RT-PCR检测结果显示,hc PGRPS3在性腺及肝胰腺中表达水平较高。经PGN或LPS刺激后,肝胰腺中hc PGRPS3表达水平显著上调,表明hc PGRPS3可能在机体抗菌免疫反应中发挥重要作用。
[Abstract]:Peptidoglycan recognition protein (PGRPs) is an important pattern recognition receptor in the innate immune system of human body. A short peptidoglycan recognition protein of Hyriopsis cumingii was cloned and expressed. In this study, the PGRPS3 gene (hc PGRPS3) fragment of Hyriopsis cumingii (Hyriopsis cumingii) obtained from high-throughput transcript group sequencing was cloned by 5-and 3-race techniques, respectively. The full-length hc PGRPS3 c-DNA sequence was obtained by splicing. The full length sequence of hc PGRPS3 c DNA was characterized by various molecular biological software. Real time fluorescence quantitative (Q RT-PCR) was used to detect its tissue distribution, and Peptidoglycan,PGN and Lipopolysaccharide, were used to detect its tissue distribution. The open reading frame of hc PGRPS3 c DNA 1438 bp, encoded 285amino acids in hepatopancreas, and the predicted molecular weight was 32.3 ku,p H 7.0. The theoretical equivalence of hc PGRPS3 c DNA was 7.98 when the molecular weight was 32.3 ku,p H 7.0. The open reading frame of hc PGRPS3 c DNA was 1438 bp, and the open reading frame of hc PGRPS3 c DNA was 858 bp,. There was no signal peptide and transmembrane structure in the sequence. The conservative analysis of amino acid sequence showed that among the other short PGRP species, Hawaii squid PGRP4 had the highest homology with hc PGRPS3 (51%). Q RT-PCR), and the expression level of hc PGRPS3 in gonads and hepatopancreas was higher. After stimulation with PGN or LPS, the expression level of hc PGRPS3 in hepatopancreas was significantly up-regulated, suggesting that hc PGRPS3 may play an important role in anti-bacterial immune response.
【作者单位】：江西师范大学生命科学学院;江西农业大学动物科学学院;
【基金】：国家自然科学基金项目(31060359) 江西省自然科学基金(20122BAB204017)资助~~
【分类号】：S917.4

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