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。AJGBP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。
β-1,3-glucan-binding protein(GBP)is a pattern recognition receptor(PRR),which can recognize pathogenic microor⁃ganisms and activate the immune response.The GBP gene(AJ-GBP)was cloned from the sequenced transcriptome of Apostichopus ja⁃ponicus 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-posi⁃tive 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 resis⁃tance to invasion of pathogenic microorganism and the actual production and utilization of AJ-GBP recombinant protein.
作者
贾昌锋
陈知雨
雷一萱
张雷
孔令明
JIA Chang-feng;CHEN Zhi-yu;LEI Yi-xuan;ZHANG Lei;KONG Ling-ming(Marine College,Shandong University,Weihai 264209,Shandong,China)
出处
《湖北农业科学》
2021年第19期135-140,共6页
Hubei Agricultural Sciences
基金
山东省自然科学基金项目(ZR2019PC027)。
