title: [Heterologous expression of attacin gene from Musca domestica in Pichia pastoris].
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.
  
  
  CONCLUSIONS
  The attacin gene from Musca domestica has been expressed in P. pastoris.
authors.affiliation: School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China.
authors.forename: Xiao-Bo
authors.initials: XB
authors.lastname: Li
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Ma
authors.forename: Jia-Yong
authors.initials: JY
authors.lastname: Zhu
authors.forename: Man-Yu
authors.initials: MY
authors.lastname: Liu
authors.forename: Xiao-Bao
authors.initials: XB
authors.lastname: Jin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006793:Houseflies / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D016133:Polymerase Chain Reaction
chemical_list: D019476:Insect Proteins; C039711:attacin antibacterial protein, insect