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Molecular Mechanisms of Self-defense system in Insect

昆虫自卫系统的分子机制

基本信息

批准号：
13660093
负责人：
MORISHIMA Isao
金额：
$ 2.24万
依托单位：
Tottori University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

1. Screening of bacteria-induced genes in Eri-silkworm, Samia cynthia riciniComparing 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.

1.蓖麻蚕（Samia cynthia ricini）细菌诱导基因的筛选用差异克隆方法比较了未感染和注射细菌的蓖麻蚕脂肪体的mRNA，分离出12个对注射细菌的蓖麻蚕脂肪体具有特异性的DNA片段，以这些DNA片段为探针筛选了免疫蓖麻蚕脂肪体的cDNA文库，获得了一种溶菌酶的cDNAs，从蓖麻链球菌免疫后的血淋巴中分离到一种溶菌酶，其N-末端氨基酸序列与其N-末端氨基酸序列一致，表明该溶菌酶具有较高的活性，并与N-末端氨基酸序列一致。该酶与其它鳞翅目溶菌酶性质进行了比较，并分离得到一种Attacin，该Attacin仅对革兰氏阴性菌有杀菌作用。从家蚕和蓖麻蚕血淋巴中分离到3个与肽聚糖（PGN）结合活性<125>不同的蛋白质，其中一个蛋白质（31 kd）为血淋巴蛋白（C-19），该蛋白质已被分离并测序，该蛋白不仅与PGN结合，而且与寡几丁质和LPS结合。第二个蛋白是促进蛋白，它是从核多角体病毒的复制中分离出来的。