在烟草毛状根表达系统中重组生产牛乳铁蛋白衍生的抗菌肽。
Recombinant production of bovine Lactoferrin-derived antimicrobial peptide in tobacco hairy roots expression system.
机构信息
Department of Plant Breeding, Faculty of Agriculture, Ilam University, Ilam, Iran.
Department of Agronomy, Faculty of Agriculture, Malayer University, Malayer, Iran.
出版信息
Plant Physiol Biochem. 2018 Feb;123:414-421. doi: 10.1016/j.plaphy.2017.12.037. Epub 2017 Dec 30.
LFchimera is a chimerical peptide containing Lactoferricin and Lactoferrampin antimicrobial peptides of bovine lactoferrin, and it has stronger bactericidal activity. Antimicrobial peptides (AMPs) like LFchimera have great potential as an alternative candidate for conventional antibiotics. Plant hairy roots provide suitable platform for fast, easy and cost-effective production of various recombinant proteins. The aim of this study was to express recombinant LFchimera in Nicotiana tabacum hairy roots and investigate its antimicrobial activity. The integration and expression of the transgene in hairy roots were confirmed by PCR and RT-PCR, respectively. LFchimera levels were quantified by ELISA and the presence of LFchimera was verified by SDS-PAGE analysis of root extracts. Biological activity of the plant derived LFchimera was confirmed by investigating the antimicrobial activity of total solution protein against Escherichia coli (ATCC 8739). Hairy root biomass reached to 4.6 g and LFchimera accumulate about 4.8 μg/g fresh weight in Erlenmeyer flasks.
Lfchimera 是一种嵌合肽，包含乳铁蛋白的乳铁肽和乳铁蛋白肽，具有更强的杀菌活性。抗菌肽（AMPs）如 Lfchimera 作为传统抗生素的替代品具有很大的潜力。植物毛状根为快速、简便和经济高效地生产各种重组蛋白提供了合适的平台。本研究旨在在烟草毛状根中表达重组 Lfchimera 并研究其抗菌活性。通过 PCR 和 RT-PCR 分别证实了转基因在毛状根中的整合和表达。通过 ELISA 定量 Lfchimera 水平，并通过 SDS-PAGE 分析根提取物来验证 Lfchimera 的存在。通过研究总溶液蛋白对大肠杆菌（ATCC 8739）的抗菌活性来证实植物来源的 Lfchimera 的生物活性。在摇瓶中，毛状根生物量达到 4.6 g，Lfchimera 积累约 4.8μg/g 鲜重。
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