Preparation and Application Analysis of a Polyclonal Antibody as Reference Protein in Helicoverpa armigera (Lepidoptera: Noctuidae)

IF 0.7 4区 农林科学 Q4 ENTOMOLOGY Journal of Entomological Science Pub Date : 2023-06-26 DOI:10.18474/JES22-71
Yun Tan, Ruoheng Jin, Zhengkun Xiao, Guo‐Hua Huang
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Abstract

Abstract A stable and specific heat shock protein 27.2 (HSP27.2) antibody was prepared and analyzed for protein level research in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). The full-length hsp27.2 was amplified from H. armigera larvae and constructed into the prokaryotic expression vector. The purified His-tag fused protein was used to immunize rabbits for the antibody preparation. Western blot analysis indicated that this antibody specifically recognized the HSP27.2 encoded by H. armigera and detected the HSP27.2 encoded by other noctuid larvae. Further analysis of HSP27.2 expression in H. armigera under infection by different pathogenic microorganisms and in different tissues showed that the expression of HSP27.2 is continually stable. The HSP27.2 antibody is efficient and capable as a reference antibody for functional studies involving genes and proteins in H. armigera and other lepidopteran insects.
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棉铃虫参考蛋白多克隆抗体的制备及应用分析
摘要制备了一种稳定特异的热休克蛋白27.2 (HSP27.2)抗体,用于棉铃虫(Helicoverpa armigera, h bner)蛋白水平研究。从棉蚜幼虫中扩增hsp27.2全长,构建原核表达载体。将纯化的His-tag融合蛋白免疫家兔制备抗体。Western blot分析表明,该抗体能特异性识别棉铃虫编码的HSP27.2,也能检测到其他夜蛾幼虫编码的HSP27.2。进一步分析HSP27.2在不同病原微生物感染下和不同组织中的表达,发现HSP27.2的表达是持续稳定的。HSP27.2抗体在棉铃虫和其他鳞翅目昆虫的基因和蛋白质的功能研究中是有效的和可作为参考抗体的。
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来源期刊
CiteScore
1.20
自引率
11.10%
发文量
40
审稿时长
>12 weeks
期刊介绍: The Journal of Entomological Science (ISSN 0749-8004) is a peer-reviewed, scholarly journal that is published quarterly (January, April, July, and October) under the auspices of the Georgia Entomological Society in concert with Allen Press (Lawrence, Kansas). Manuscripts deemed acceptable for publication in the Journal report original research with insects and related arthropods or literature reviews offering foundations to innovative directions in entomological research
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