Purification and cDNA cloning of inducible antibacterial peptides fromProtaetia brevitarsis (Coleoptera)
| Autor: | In Hee Lee, Hye Suk Yoon, Sang Yong Lee, Hak R. Kim, Chung Sik Choi, Sung Moon Yeo, Chang Seok Lee |
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| Rok vydání: | 2003 |
| Předmět: |
DNA
Complementary Physiology Molecular Sequence Data Microbial Sensitivity Tests Gram-Positive Bacteria medicine.disease_cause Biochemistry Hemolymph Gram-Negative Bacteria medicine Animals Amino Acid Sequence Cloning Molecular Escherichia coli Peptide sequence Base Sequence biology Molecular mass Reverse Transcriptase Polymerase Chain Reaction Fast protein liquid chromatography General Medicine Blotting Northern biology.organism_classification Molecular biology Anti-Bacterial Agents Coleoptera Reverse transcription polymerase chain reaction Larva Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Insect Science Insect Proteins Electrophoresis Polyacrylamide Gel Primer (molecular biology) Peptides Bacteria |
| Zdroj: | Archives of Insect Biochemistry and Physiology. 52:92-103 |
| ISSN: | 1520-6327 0739-4462 |
| DOI: | 10.1002/arch.10072 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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