Purification and cDNA cloning of inducible antibacterial peptides from Protaetia brevitarsis (Coleoptera).
Arch Insect Biochem Physiol
; 52(2): 92-103, 2003 Feb.
Article
en En
| MEDLINE
| ID: mdl-12529864
Three antibacterial peptides, named protaetins 1, 2, and 3, were purified and characterized from immunized larval hemolymph of Protaetia brevitarsis, a fruit tree pest in Korea. Also, protaetin 1 was cloned. Acid extraction, gel filtration, preparative acid-urea PAGE, and reversed-phase FPLC were used for purification of peptides. Protaetins 1 and 3 had molecular masses of 7.5 and 12 kDa on Tricine SDS-PAGE, respectively, and the molecular mass of protaetin 2 was 9,283.95 Da as determined by MALDI-TOF mass spectrometry. In an antibacterial assay, protaetins showed antibacterial activities against a panel of Gram-positive and -negative bacteria. For the RT-PCR (reverse transcription polymerase chain reaction) to obtain the complete primary sequence, the primer was designed according to the N-terminal amino acid sequence of protaetin 1. Amino acid sequence homology of protaetin 1 with holotricin 2, an antibacterial peptide from Holotrichia diomphalia, showed 99% identity. Northern blot analysis showed that the protaetin 1 gene was strongly expressed in the fat body after Escherichia coli injection, but not in normal fat body. Also, it was expressed in the gut, but was much weaker after immunization.
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Base de datos:
MEDLINE
Asunto principal:
Péptidos
/
Escarabajos
/
ADN Complementario
/
Proteínas de Insectos
/
Antibacterianos
Idioma:
En
Revista:
Arch Insect Biochem Physiol
Asunto de la revista:
BIOLOGIA
/
BIOQUIMICA
Año:
2003
Tipo del documento:
Article