摘要
[目的]获得抗菌肽LL-1的完整基因序列,并研究其生物学特性及抗菌效果,为进一步研究开发抗菌肽药物奠定基础。[方法]本研究通过桃蛀螟转录组测序和比对分析发现一种抗菌肽,命名为LL-1。通过RACE扩增技术获得其完整基因序列,从而推导其氨基酸序列。通过多重序列比对,获得其裂解后的成熟多肽并进行化学合成。通过建立大肠杆菌感染鸡肉模型,评价LL-1的抑菌效果,并检测其耐盐和耐高温特性。[结果]LL-1基因全长474 bp, ORF长192 bp,编码63个氨基酸残基,成熟的LL-1含有39个氨基酸残基,为天蚕素抗菌肽。50 mg/kg的LL-1能极显著抑制大肠杆菌在鸡肉中增殖(P<0.01)。耐温度特性分析结果表明,在4~50℃范围内,温度对抗菌肽LL-1的抑菌效果没有显著影响;与37℃相比,当100℃处理1 h后,抗菌肽LL-1的抑菌活性显著降低(P<0.05)。耐盐特性分析结果表明,在0~200 mmol/L NaCl溶液处理后,抗菌肽LL-1的抑菌效果未出现显著变化(P>0.05)。[结论]抗菌肽LL-1具有较好的稳定性及抗菌活性,具有开发成新型药物的潜力。
[Objective]The aim of this study was to obtain the complete gene sequence of antimicrobial peptide LL-1 and study its biological characteristics and antibacterial effect,in order to lay a foundation for further research and development of antimicrobial peptide drugs.[Methed]In this study,an antimicrobial peptide,named LL-1,was discovered through transcriptome sequencing and sequence alignment analysis of Conogethes punctiferalis.The complete gene sequence was obtained by RACE technology,and its corresponding amino acid sequence was derived.After cleavage,the mature peptide sequence was analyzed using multiple sequence alignment and chemically synthesized.Subsequently,the antibacterial effect of antimicrobial peptide LL-1 was evaluated by establishing a model of E.coli infection in chicken meat.Finally,the salt and temperature resistance of antimicrobial peptide LL-1 were analyzed.[Result]The results showed that the LL-1 gene had a total length of 474 bp with an ORF length of 192 bp encoding 63 amino acids.Mature antimicrobial peptide LL-1 consisted 39 amino acid residues,and belonged to cecropin family.At the concentration of 50 mg/kg,antimicrobial peptide LL-1 could extremely significantly inhibit E.coli growth in chicken meat(P<0.01).The temperature tolerance analysis showed that the antibacterial effect of antimicrobial peptide LL-1 was not significantly affected by temperature within the range of 4-50℃.After being treated at 100℃for 1 h,the antibacterial activity of antimicrobial peptide LL-1 was significantly lower than that of antimicrobial peptide LL-1 treated at 37℃(P<0.05).The salt tolerance analysis showed that the antibacterial effect of antimicrobial peptide LL-1 did not show a significant change after being treated with NaCl solutions of 0-200 mmol/L concentration.[Conclusion]Antimicrobial peptide LL-1 had good stability and antibacterial activity,and had the potential to be developed into new drugs.
作者
周玲玲
王宇航
李亦菲
柴永乐
张明亮
张元臣
连凯琪
ZHOU Lingling;WANG Yuhang;LI Yifei;CHAI Yongle;ZHANG Mingliang;ZHANG Yuanchen;LIAN Kaiqi(School of Biotechnology and Food Science,Anyang Institute of Technology,Anyang 455000,China;Henan Joint International Research Laboratory of Veterinary Biologics Research and Application,Anyang 455000,China;Taihang Mountain Forest Pests Observation and Research Station of Henan Province,Linzhou 456550,China)
出处
《中国畜牧兽医》
北大核心
2025年第1期491-497,共7页
China Animal Husbandry & Veterinary Medicine
基金
河南省科技攻关计划项目(242102110073)
安阳工学院博士启动基金项目(BSJ2024005)
安阳市重点研发与推广专项(2023C01NY013)。
关键词
抗菌肽
化学合成
生物学特性
抗菌效果
序列比对
antibacterial peptides
chemical synthesis
biological characteristics
antibacterial effect
sequence alignment
作者简介
周玲玲,E-mail:13700719135@163.com;通信作者:张元臣,E-mail:zhangyuanchen0110@163.com;通信作者:连凯琪,E-mail:liankaiqi616@163.com。
