1995 Fiscal Year Final Research Report Summary
Use of Bombyx mori Mutants to Study on Insect Biochemistry
| Project/Area Number |
06044175
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
FUJII Hiroshi FACULTY OF AGRICULTURE・KYUSHU UNIVERSITY・ASSOCIATE PROFESSOR, 農学部, 助教授 (10038268)
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| Co-Investigator(Kenkyū-buntansha) |
LAW Jhon h ARIZONA UNIVERSITY PROFESSOR, 教授
WELLS Michael a ARIZONA UNIVERSITY・PROFESSOR, 教授
TSUCHIDA Kozo NATIONAL INSTITUTE HEALTH・RESEARCHER, 主任研究官 (40231435)
MAEKAWA Hideaki NATIONAL INSTITUTE HEALT・HEAD, 室長 (60100096)
SUMITA Motoyuki KYOTO INSTITUTE TECHNOLOGY・ASSOCIATE PROFESSOR, 繊維学部, 助教授 (50127164)
KOBAYASHI Masahiko TOKYO UNIVERSITY・PROFESSOR, 農学部, 教授 (60162020)
SUZUKI Akinori TOKYO UNIVERISITY・PROFESSOR, 農学部, 教授 (90011907)
DOIRA Hiroshi KYUSHU UNIVERSITY・PROFESSOR, 農学部, 教授 (30038210)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Bombyx mori / CBP / YI and Y+^I midgut / Cnad+^C silkgland / LTP / membrane permeability / immunocytology / immuno-gold / electron microscop |
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| Research Abstract |
Wells lab isolated lutein bound protein (40kDa) from the midgut of silkworm. Fuji also isolated and characterized carotinoid binding proitein (600kDa) from the midgut of the Y+^I silkworm. This protein is identical with CBP in hemolymph on the bisis of immunological similarity and the same biochemical properties as to CBP.The midgut of the YI silkworm contained a yellow colored protein, but did not in the Y+^I silkworm. The color pigment was carotinoid. But the protein did not react with anti-CBP antibody. Hence it is a new carotinoid binding protein other than CBP.
cDNA library from the midgut of the Y+^I silkworm was constracted. cDNA encoding CBP has been cloned by using anti-CBP antibody and several clones were obtained. Two clones were analyzed nucleotide sequence, but they were not identical to CBP.
Immunocytochemical analysis of carotinoid binding protein (CBP) in the midgut and middle silkgland of silkworm, Bombyx mori. Immunogold technique showed that CBP was located in rough ER,and in the small vesicle associated with smooth ER.Carotinoid was secreated into hemolymph from midgut as carotinoid bound CBP complex by exocytosis, incorporated into the middle part of silkgland from hemolymph as the complex and finally tansferrd to sericin prior to excretion of sericine particles.
A lipid transfer particles (LTP) has been purified from the hemolymph of Bombyx mori. the LTP has three apoproteins (79% of particles mass) apo LTP-I,apoLTP=IInd apoLTP=III,Mr=350,000,85,000and 60,000. The lipid of LTP is phosphalipid (54%), monoacylgicerol (8.3%), diaacylglycerol (17%), free fatty acid (14.3%), triacylglycerol (5.9%) and hydrocarbon (0.4%).This protein is able to catalyze the transfer of a number of lipids among different lipid particle.
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