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  • Title: [Heterologous expression of attacin gene from Musca domestica in Pichia pastoris].
    Author: Li XB, Wang TT, Ma Y, Zhu JY, Liu MY, Jin XB.
    Journal: Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi; 2010 Oct 30; 28(5):332-6. PubMed ID: 21351542.
    Abstract:
    OBJECTIVE: To construct recombinant expression plasmid with an antibacterial peptide (attacin) mature peptide gene from Musca domestica and express an antibacterial peptide attacin in Pichia pastoris. METHODS: The coding region of attacin mature peptide was cloned into the expression vector pPIC9K. The recombinant plasmid was linearized by digestion with Sac I and transformed into P. pastoris GS115 by electroporation. The insert clones containing multiple copies were selected with geneticin, pha, enotype and PCR. The expression of P. pastoris GS115/pPIC9K-attacin were induced with methanol. The supernatant of the expressed protein was analyzed by SDS-PAGE and Western blotting. The anti-bacterial activity of expression product was analyzed by agarose diffusion assay. RESULTS: The expression plasmid was constructed and transformed into P. pastoris GS115. SDS-PAGE and Western blotting analysis indicated that the recombinant containing recombinant plasmid pPIC9K-attacin expressed a Mr 22000 protein. The expression product inhibited the growth of E. coli K12D31. CONCLUSION: The attacin gene from Musca domestica has been expressed in P. pastoris.
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