| 新型欧洲鳗鲡抗菌肽elecilin在毕赤酵母中的表达 |
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| 引用本文： | 张丽娟,吴唯维,李素一,陈华,柯翎,陈叙,林晨韬.新型欧洲鳗鲡抗菌肽elecilin在毕赤酵母中的表达[J].福建畜牧兽医,2021(2):19-21. |
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| 作者姓名： | 张丽娟 吴唯维 李素一 陈华 柯翎 陈叙 林晨韬 |
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| 作者单位： | 福建省农业科学院生物技术研究所;福建师范大学生命科学学院;福建师范大学南方海洋研究院福建省微藻种质改良工程技术研究中心;福建师范大学福建省特色海洋生物资源可持续利用重点实验室 |
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| 基金项目： | 福建省科技计划公益类专项(2017R1019-9);福建省农业科学院青年人才创新基金(YC2017-5);福建省农业科学院科技创新团队项目(STIT2017-3-1)资助。 |
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| 摘 要： | 前期采用差减杂交的方法筛选到新型欧洲鳗鲡抗菌肽elecilin,其基因全长为477 bp,编码159个氨基酸,理论分子量约17.4 kDa。为了获得具活性的大量表达的抗菌肽,本研究利用毕赤酵母表达了elecilin。首先将elecilin基因克隆至酵母分泌表达载体pPICZaA,构建了重组分泌表达质粒pPICZaA-elecilin,电击转化毕赤酵母X-33。经Zeocin筛选和PCR鉴定,获得重组酵母菌,通过甲醇诱导,SDS-PAGE和Western blot鉴定,证实诱导后的培养基中存在与预期分子量大小相一致的蛋白,能被His-tag单克隆抗体识别,表明利用酵母成功表达了欧洲鳗鲡抗菌肽elecilin。这为进一步研究该抗菌肽的功能及作为饲料添加剂的应用奠定了基础。
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| 关 键 词： | 欧洲鳗鲡 抗菌肽 表达 毕赤酵母 |
Expression of a novel Anguilla anguilla antimicrobial peptide,elecilin,in Pichia pastoris |
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| Zhang Lijuan,Wu Weiwei,Li Suyi,Chen Hua,Ke Ling,Chen Xu,Lin Chentao.Expression of a novel Anguilla anguilla antimicrobial peptide,elecilin,in Pichia pastoris[J].Fujian Journal of Animal Husbandry and Veterinary,2021(2):19-21. |
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| Authors: | Zhang Lijuan Wu Weiwei Li Suyi Chen Hua Ke Ling Chen Xu Lin Chentao |
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| Institution: | (Biotechnology Institute,Fujian Academy of Agricultural Sciences,Fuzhou 350003;College of Life Sciences,Fujian Normal University,Fuzhou 350007;Center of Engineering Technology Research for Microalgae Germplasm Improvement of Fujian,Southern Institute of Oceanography,Fujian Normal University,Fuzhou 350117;Fujian Key Laboratory of Special Marine Bio-resources Sustainable Utilization,Fujian Normal University,Fuzhou 350117) |
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| Abstract: | A novel antimicrobial peptide,elecilin,was obtained by subtractive hybridization from Anguilla anguilla.The gene is 477 bp in length,encoding 159 amino acids,with a theoretical molecular weight of about 17.4 kDa.In order to obtain highly expressed bioactive antimicrobial peptides,Pichia pastoris was used to express the gene,elecilin.Firstly,the gene was cloned into the yeast secretion expression vector pPICZaA to construct plasmid pPICZaA-elecilin,then it was transformed into Pichia pastoris X-33 by electroporation and recombinant yeast was obtained by Zeocin screening and PCR identification.After methanol induction,SDS-PAGE and Western blot analysis indicated that a specific band with similar molecular weight as expected size was found,and it could be recognized by anti-His-tag monoclonal antibody,which indicated that elecilin was successfully expressed by Pichia pastoris.This results lay a foundation for further research on the antimicrobial peptide function and its application as feed additive. |
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| Keywords: | Anguilla anguilla Antimicrobial peptide Expression Pichia pastoris |
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