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Molecular cloning of a novel cecropin-like peptide gene from the swallowtail butterfly, Papilio xuthus

Kim, Seong-Ryul;Choi, Kwang-Ho;Kim, Sung-Wan;Hwang, Jae-Sam;Goo, Tae-Won;Kim, Iksoo

  • Received : 2015.10.30
  • Accepted : 2015.11.03
  • Published : 2015.12.31

Abstract

A new cecropin-like antimicrobial peptide (Px-CLP) gene was isolated from the immunechallenged larvae of the swallowtail butterfly, Papilio xuthus, by employing annealing control primer (ACP)-based GeneFishing PCR. The full-length cDNA of Px-CLP is 310 nucleotides encoding a 70 amino acid precursor that contains a putative 22-residue signal peptide, a 4-residue propeptide, a presumed 37-residue mature peptide, and an uncommon 7-residue acidic pro-region at the C-terminus. The deduced amino acid sequence of Px-CLP showed significant identities with other Lepidopteran cecropin D type peptides. RT-PCR revealed that the Px-CLP transcript was detected at significant level after injection with bacterial lipopolysaccharide (LPS). The peptides with or without C-terminal acidic sequence region were synthesized on-solid phage and submitted to antibacterial activity assay. The synthetic 37-mer peptide (Px-CLPa), which removed C-terminal acidic sequence region, was showed exclusively antibacterial activity against E. coli ML35; meanwhile, a 44-mer peptide (Px-CLPb) with C-terminal acidic peptide region was not active. This result suggests that Px-CLP is produced as a larger precursor containing a C-terminal pro-region that is subsequently removed by C-terminal modification.

Keywords

Papilio xuthus;antimicrobial peptide;RT-PCR;antibacterial activity

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Acknowledgement

Grant : 누에 생체공장화 시스템 이용 천연항생제 생산 기술