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从黑缘红萤中克隆和功能表征一个新型β-GRP 基因。

Cloning and Functional Characterization of a Novel β-GRP Gene From Melanotus cribricollis.

机构信息

Zhejiang Academy of Forestry, Hangzhou 310023, China.

Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Hangzhou 311400, China.

出版信息

J Insect Sci. 2022 Sep 1;22(5). doi: 10.1093/jisesa/ieac051.

Abstract

In this study, a novel β-1,3-glucan recognition protein gene (β-GRP) was identified from Melanotus cribricollis, and its potential role in the immune response was investigated. The full length of β-GRP cDNA (Accession Number: MT941530) was 1644 bp, encoding a protein composed of 428 amino acids. The theoretical molecular weight and the isoelectric point were 51.53 kDa and 6.17, respectively. The amino acid sequence of β-GRP from M. cribricollis was closely related to that of. β-GRP-like from Photinus pyralis, and was predicted to contain conserved GH16 domain without glucanase active site. The results of real-time quantitative PCR showed that fungal infection of Metarhizium pingshaense may significantly upregulated the expression level of β-GRP gene. The RNAi suppression of β-GRP gene expression significantly increased the corrected cumulative mortality. Meanwhile, antimicrobial peptide genes defensin and lysozyme were significantly downregulated after interference of β-GRP. Taken together, these results suggest that β-GRP of M. cribricollis probably participates in the host immune system by mediating the expression of antimicrobial peptides. This study provides comprehensive insights into the innate immune system of insect larvae.

摘要

在这项研究中,从墨天牛中鉴定出一种新型的β-1,3-葡聚糖识别蛋白基因(β-GRP),并研究了其在免疫反应中的潜在作用。β-GRP cDNA 的全长(注册号:MT941530)为 1644 bp,编码由 428 个氨基酸组成的蛋白质。理论分子量和等电点分别为 51.53 kDa 和 6.17。来自墨天牛的β-GRP 氨基酸序列与 Photinus pyralis 的β-GRP-like 密切相关,预测其含有保守的 GH16 结构域,而没有葡聚糖酶活性位点。实时定量 PCR 的结果表明,绿僵菌感染米象可能会显著上调β-GRP 基因的表达水平。β-GRP 基因表达的 RNAi 抑制显著增加了校正后的累积死亡率。同时,β-GRP 干扰后防御素和溶菌酶等抗菌肽基因的表达显著下调。总之,这些结果表明,墨天牛的β-GRP 可能通过调节抗菌肽的表达参与宿主免疫系统。本研究为昆虫幼虫的先天免疫系统提供了全面的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5b0/9459437/0120758a9a11/ieac051_fig7.jpg

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