仿刺参葡聚糖结合蛋白的重组表达与功能研究

doi:10.14088/j.cnki.issn0439-8114.2021.19.029

湖北农业科学 ›› 2021, Vol. 60 ›› Issue (19): 135-140.doi: 10.14088/j.cnki.issn0439-8114.2021.19.029

仿刺参葡聚糖结合蛋白的重组表达与功能研究

贾昌锋, 陈知雨, 雷一萱, 张雷, 孔令明

山东大学海洋学院,山东威海 264209

收稿日期:2021-02-11 出版日期:2021-10-10 发布日期:2021-10-22
作者简介:贾昌锋（1999-）,男,新疆乌鲁木齐人,在读本科生,研究方向为海水养殖与资源利用,（电话）19819667268（电子信箱）jia9829@outlook.com。
基金资助:
山东省自然科学基金项目（ZR2019PC027）

Recombinant expression and functional study of dextran binding protein in Apostichopus japonicus

JIA Chang-feng, CHEN Zhi-yu, LEI Yi-xuan, ZHANG Lei, KONG Ling-ming

Marine College, Shandong University,Weihai 264209,Shandong,China

Received:2021-02-11 Online:2021-10-10 Published:2021-10-22

摘要/Abstract

摘要： β-1,3-葡聚糖结合蛋白（β-1,3-glucan-binding protein,GBP）是一种模式识别受体（Pattern recognition receptor,PRR）,可识别病原微生物从而激活机体免疫应答。基于聚合酶链反应（Polymerase Chain Reaction,PCR）从已测序的仿刺参（Apostichopus japonicus）转录组中克隆了仿刺参葡聚糖结合蛋白基因（AJ-GBP）。核苷酸序列分析表明,该基因全长660 bp,可编码219个氨基酸,不具有信号肽。AJ-GBP重组蛋白理论分子质量为24.64 kDa,等电点为5.94,属于糖基水解酶家族16,且在第17位和第193位存在糖基化位点。试验结果表明,该重组蛋白对革兰氏阳性菌、革兰氏阴性菌具有凝集、结合和抑制作用;对葡聚糖具有明显结合活性,表明AJ-GBP重组蛋白可能参与了仿刺参的先天免疫反应中,研究为了解仿刺参抵抗病原微生物入侵的机制以及AJ-GBP重组蛋白实际生产利用奠定了理论基础。

关键词: 仿刺参（Apostichopus japonicus）, β-1, 3-葡聚糖结合蛋白, 重组表达, 功能分析

Abstract: β-1,3-glucan-binding protein （GBP） is a pattern recognition receptor （PRR）, which can recognize pathogenic microorganisms and activate the immune response. The GBP gene （AJ-GBP） was cloned from the sequenced transcriptome of Apostichopus japonicus by Polymerase Chain Reaction （PCR）. Nucleotide sequence analysis showed that the total length of the gene was 660 bp, and it could encode 219 amino acids without signal peptide. The theoretical molecular weight of AJ-GBP protein was 24.64 kDa, and the isoelectric point was 5.94. It belonged to the glycosyl hydrolases family 16, and there were glycosylation sites at positions 17 and 193. The results of protein activity test showed that the recombinant protein had agglutination, binding and inhibition effects on Gram-positive and Gram-negative bacteria. It has obvious binding activity to glucan. These results suggest that AJ-GBP may be involved in the innate immune response of Apostichopus japonicus. This study laid a theoretical foundation for understanding the mechanism of resistance to invasion of pathogenic microorganism and the actual production and utilization of AJ-GBP recombinant protein.

Key words: Apostichopus japonicus, β-1, 3-glucan-binding protein, recombinant expression, functional analysis

中图分类号:

Q819

贾昌锋, 陈知雨, 雷一萱, 张雷, 孔令明. 仿刺参葡聚糖结合蛋白的重组表达与功能研究[J]. 湖北农业科学, 2021, 60(19): 135-140.

JIA Chang-feng, CHEN Zhi-yu, LEI Yi-xuan, ZHANG Lei, KONG Ling-ming. Recombinant expression and functional study of dextran binding protein in Apostichopus japonicus[J]. HUBEI AGRICULTURAL SCIENCES, 2021, 60(19): 135-140.

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仿刺参葡聚糖结合蛋白的重组表达与功能研究

Recombinant expression and functional study of dextran binding protein in Apostichopus japonicus

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