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  1. National Taiwan Ocean University Research Hub

Application of Antimicrobial Peptides in Aquaculture

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基本資料

Project title
Application of Antimicrobial Peptides in Aquaculture
Code/計畫編號
92農科-4.2.3-漁-F1(9)
Translated Name/計畫中文名
抗菌peptide在水產養殖之應用研究
 
Project Coordinator/計畫主持人
Cheng-Hui Lin
Funding Organization/主管機關
Council of Agriculture,Executive Yuan
 
Department/Unit
Department of Aquaculture
Website
https://www.grb.gov.tw/search/planDetail?id=960819
Year
2003
 
Start date/計畫起
01-01-2003
Expected Completion/計畫迄
31-12-2003
 
Bugetid/研究經費
650千元
 
ResearchField/研究領域
生物技術(農)
漁業
 

Description

Abstract
我們已利用化學合成或購得數種抗菌peptide,並利用輻射擴散法分析,測得抗菌peptideKM之抗菌活性最好,且針對抗菌peptideKM設計primer,並完成構築抗菌peptideKM表現質體.為了避免表現出來的重組抗菌peptide會對宿主細胞造成毒害,在抗菌peptideN端前加入一段分子量較大的蛋白質,以期表現出來的融合蛋白能中和重組抗菌peptideKM對宿主細胞所造成的毒害.本實驗所使用融合蛋白基因為Thioredoxin protein部分片段,包含109個胺基酸,此蛋白亦可增進融合蛋白在細胞質(cytoplasm)的可溶性.重組抗菌peptide經IPTG誘導產生後,再以His-Taq column純化,經由Enterokinase剪切後,並利用輻射擴散法分析,測得重組抗菌peptide具有抗菌活性.本年度我們將利用細菌製造之重組抗菌peptide,測試重組抗菌peptide對各種水產生物病原菌的抗菌活性. The antimicrobial peptides were synthesized by chemical method or purchased and analyzed by radial diffusion assay. The results show that the antimicrobial activity of antimicrobial peptide KM7K is the best. The primers encoded antimicrobial peptide were designed, chemically synthesized, and inserted into expression vector. In order to avoid destroying host cells, a fragment of thioredoxin protein is added into the N-terminal of recombinant antimicrobial peptide. The fusion protein becomes soluble and is produced in the cytoplasm of host cells. The recombinant antimicrobial peptide is induced by IPTG, purified by His-Taq column, digested by enterokinase, and analyzed by radial diffusion assay. We found that the recombinant antimicrobial peptide contains the antimicrobial activity. In the plan of this year, we try to produce large scale of recombinant antimicrobial peptide and examine the antimicrobial activity of recombinant antimicrobial peptide to against the aquatic pathogens.
 
 
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