Application of Antimicrobial Peptides in Aquaculture

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Project title

Code/計畫編號

91農科-3.1.1-漁-F1(12)

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=941277

Year

2002

Start date/計畫起

01-01-2002

Expected Completion/計畫迄

31-12-2002

Bugetid/研究經費

622千元

ResearchField/研究領域

生物技術(農)
漁業

上年度已利用化學合成抗菌peptide，並利用輻射擴散法分析，測得抗菌peptide KM 之抗菌活性最好。且針對抗菌peptide KM 設計primer，並完成構築抗菌peptide KM表現載體，得知由細菌產生之重組抗菌peptide KM具有抗菌活性。為了避免表現出來的重組抗菌peptide KM會對宿主細胞造成毒害，在KM基因的上游插入一段分子量較大、且表現出的蛋白質傾向較為可溶性的基因片段，以期表現出來的融合蛋白能中和重組抗菌peptides KM對宿主細胞所造成的毒害，以及使融合蛋白更為可溶性，以方便純化。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，可增進表現融合蛋白在細胞質（cytoplasm）的可溶性。重組抗菌peptides以IPTG誘導後，再以His-Taq column純化，並分析其抗菌活性。 The antimicrobial peptides were synthesized by chemical method 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, and analyzed by radial diffusion assay.

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Application of Antimicrobial Peptides in Aquaculture

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