埃及伊蚊肽聚糖识别蛋白PGRP-S2的克隆及原核表达  

CLONING AND PROKARYOTIC EXPRESSION OF AEDES AEGYPTI PEPTIDOGLYCAN RECOGNITION PROTEIN PGRP-S2

作　　者：蔡瑜婷 杨小祯 陈春梅 王焌翔 余思宸 黄恩炯 张灵玲[1,2] 关雄[1] CAI Yu-ting;YANG Xiao-zhen;CHEN Chun-mei;WANG Jun-xiang;YU Si-chen;HUANG En-jiong;ZHANG Ling-ling;GUAN Xiong(Key Laboratory of Biopesticide and Chemical Biology,Ministry of Education,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;Department of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;Fujian Medical University,Fuzhou 362002,Fujian,China;Fuzhou International Healthcare Center,Fuzhou 350001,Fujian,China)

机构地区：[1]福建农林大学生物农药与化学生物学教育部重点实验室,福州350002 [2]福建农林大学生命科学学院,福州350002 [3]福建医科大学公共卫生学院,福州350122 [4]福州国际旅行卫生保健中心,福州350001

出　　处：《寄生虫与医学昆虫学报》2022年第2期91-97,共7页Acta Parasitologica et Medica Entomologica Sinica

基　　金：福建农林大学创新基金(CXZX2020124B);生物农药与化学生物学教育部重点实验室开放课题(Keylab2018-04);闽台合作有害生物生态防控国家重点实验室开放课题(SKL2019007);害虫绿色防控福建省高等学校重点实验室开放研究基金(PTJH1700701)。

摘　　要：昆虫可通过体内肽聚糖识别蛋白识别外来微生物的肽聚糖而启动免疫机制,从而降低微生物对昆虫自身的影响,但关于蚊虫的相关报道较少。为了解埃及伊蚊的肽聚糖识别蛋白特性,本研究通过分子克隆技术,成功克隆并测序获得了埃及伊蚊肽聚糖识别蛋白AePGRP-S2的ORF序列。该ORF序列有510个碱基,可编码170个氨基酸。生信分析发现其具有典型的PGRP超家族保守结构域与酰胺酶结构域,表明该基因序列很有可能为埃及伊蚊的肽聚糖识别蛋白基因,且可能具有直接的抗菌活性。通过系统进化树分析,发现埃及伊蚊PGRP-S2与埃及伊蚊的PGRP-S的亲缘关系最近;为进一步获得埃及伊蚊肽聚糖识别蛋白AePGRP-S2,本研究构建表达载体,并将靶基因原核表达于大肠杆菌BL21(DE3)中,通过SDS-PAGE及Western Blot的检测,获得46 kDa目的蛋白,为进一步深入了解埃及伊蚊体内肽聚糖识别蛋白PGRP的功能及作用机制奠定基础。Insects can recognize the peptidoglycan of foreign microorganisms through peptidoglycan recognition proteins in vivo and activate the immune mechanism,thus reducing the impact of microorganisms on insects.However,there are limited reports about mosquitoes.In order to character Aedes aegypti peptidoglycan recognition protein,AePGRP-S2 was cloned and sequenced successfully by molecular cloning technology.The open reading frame of the PGRP sequence was 510 nt,encoding 170 amino acids.Phylogenetic tree analysis showed that Ae.aegypti PGRP-S2 was the most closely related to PGRP-S of Ae.aegypti.To further obtain AePGRP-S2,the expression vector was constructed and the target gene was prokaryotic expressed in Escherichia coli BL21(DE3).And,the 46 kDa target protein was finally detected by SDS-PAGE and Western Blot analysis.All of these provide the basis for further understanding the function and mechanism of peptidoglycan recognition protein PGRP in Ae.aegypti.

关 键 词：埃及伊蚊 肽聚糖识别蛋白 细胞免疫 克隆 表达 

分 类 号：R184.31[医药卫生—流行病学]