2002 Fiscal Year Final Research Report Summary
Molecular Mechanisms of Self-defense system in Insect
| Project/Area Number |
13660093
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | Tottori University |
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Principal Investigator |
MORISHIMA Isao Tottori University, Faculty of Agriculture,Professor, 農学部, 教授 (30032296)
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| Co-Investigator(Kenkyū-buntansha) |
YAMANO Yoshiaki Tottori University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (00182593)
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| Project Period (FY) |
2001 – 2002
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| Keywords | insect immunity / Antibacterial protein / Peptidglycan recognition protein / Attacin / Lysozyme / Samia cynthia ricini / Bombyx mori |
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| Research Abstract |
1. Screening of bacteria-induced genes in Eri-silkworm, Samia cynthia ricini
Comparing mRNAs in the fat bodies from naive and bacteria-injected larvae by means differential cloning method,12 DNA fragments,which were specific for bacteria-injected larvae,were isolated.After screened a cDNA library of immunized Samia fat body with these DNA fragments as probes,cDNAs for a lysozyme,three attacins and two peptidoglycan recognition proteins(PGRP)have been cloned and sequenced.
2.Isolation and characterization of lysozyme and attacin.
A lysozyme was isolated from immunized hemolymph of S.ricini.The N-terminal amino acid sequence was in perfect match of that deduced from the cDNA.The properties of the enzyme was compared with other lepidopteran lysozyme.An attacin was similarly isolated and characterized.The attacin was bactericidal only to Gram-negatives.
3.Isolation and characterization of peptidoglycan-binding proteins from Bombyx mori and S.ricini.
Detecting peptidoglycan (PGN) binding activity by means of photo affinity labeling with ^<125>I-PGN,three proteins,which were different from PGRP,were isolated and characterized from the hemolymph of B.mori.One of the proteins (31kd) was the hemolymph protein (C-19),which had been isolated and sequenced,but whose physiological role was not known.The protein binds not only to PGN but oligo chitin and LPS. The second protein was promoting protein,which had been isolated as the protein stimulated replication of nucleopolyhedrovirus.Two PGRP were isolated from immunized hemolymph of Samia larvae.
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