亚洲玉米螟肽聚糖识别蛋白OfPGRP-SA的表达与功能分析

Expression and functional analysis of Ostrinia furnacalis peptidoglycan recognition proteins OfPGRP-SA

目的:明确肽聚糖识别蛋白PGRP在亚洲玉米螟先天免疫中的功能。方法:构建质粒表达载体,进行重组蛋白表达、纯化及Western blot鉴定,对获得的OfPGRP-SA重组蛋白进行抗菌活性、细菌凝集、酰胺酶活性、酚氧化酶反应和黑化反应等体外试验。结果:通过SDS-PAGE和Western blot鉴定,OfPGRP-SA蛋白纯化后条带单一,与预期大小一致。对重组蛋白体外活性分析表明,OfPGRP-SA对革兰氏阳性菌有较强的诱导作用,说明OfPGRP-SA可能参与Toll途径的激活。结论:进一步明确OfPGRP-SA在亚洲玉米螟先天免疫应答中的作用,有助于更好地利用病原微生物对亚洲玉米螟进行生物防治。

Objective:To clarify the function of peptidoglycan recognition protein PGRP in the innate immunity of Ostrinia furnacalis.Methods:Plasmid expression vectors were first constructed,followed by recombinant protein expression,purification and Western blot identification,and in vitro experiments on the obtained recombinant proteins were performed for antibacterial activity,bacterial agglutination,amidase activity,phenol oxidase reaction and blackening reaction.Results:The results of identification by SDS-PAGE and Western blot showed that the OfPGRP-SA protein obtained by expression had a single band after purification and was consistent with the expected size.The in vitro activity analysis of the recombinant protein showed that OfPGRP-SA had a strong induction effect on Gram-positive bacteria,indicating that OfPGRP-SA may be involved in the activation of Toll pathway.Conclusion:The role of OfPGRP-SA in the innate immune response of Ostrinia furnacalis was further clarified by the experiment,which helps to better utilize pathogenic microorganisms for biological control of Ostrinia furnacalis.