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- W2351001891 abstract "Objective:To express Myticin A in the methylotrophic yeast Pichia pastoris and to test the bioactivity of the recombinant peptide in vitro.Methods: Total RNA was isolated from adult mussel body fluid and mRNA was used to construct a cDNA library.Reverse transcription PCR was used to obtained Myticin A gene fragment.A recombinant plasmid of pMD18-Myticin A was constructed.The Myticin A gene was inserted into the vector pPIC9K and the product was transformed into strain GS115.The transformants were selected and the recombinant peptide was expressed.The expressed sample was analyzed by Tricine-SDS-PAGE and purified by CM-sepharose and Sephadex G25 column chromatography.The biological activities were also assessed.Results:The pPIC9K-Myticin A was obtained and the stable recombinant Pichia pastoris strains were selected.Myticin A was successfully expressed as identified by Tricine-SDS-PAGE.The recombinant Myticin A was firstly purified by CM-sepharose column chromatography and the active fraction was collected and further purified by Sephadex G25 with the maximal purity reaching 90%.Purified sample was analyzed by Tricine-SDS-PAGE.Myticin A showed a strong inhibitory effect against the growth of gram-positive bacteria,specially to the Bacillus megaterium.It also inhibited the growth of tumor cell lines including SW1990,L929 and LoVo.Conclusion:An antimicrobial peptide,Myticin A,has been cloned from the body fluid of mussel and expressed in Pichia pastoris.The recombinant Myticin A exhibits a strong inhibitory effects on the growth of gram-positive bacteria, especially on Bacillus megaterium." @default.
- W2351001891 created "2016-06-24" @default.
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- W2351001891 date "2005-01-01" @default.
- W2351001891 modified "2023-09-25" @default.
- W2351001891 title "Cloning,expression and biological characterization of Myticin A,an antimicrobial peptide from mussel(Mytilus)" @default.
- W2351001891 hasPublicationYear "2005" @default.
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