1990 Fiscal Year Final Research Report Summary
Regulation of Carotenoid Permeability by Gene and Analysis its Structure
| Project/Area Number |
63480049
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
蚕糸学
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| Research Institution | Kyushu University |
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Principal Investigator |
FUJII Hiroshi Kyushu University, Faculty of Agriculture. Associate Professor, 農学部, 助教授 (10038268)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAGUCHI Yutaka Kyushu University, Faculty of Agriculture. Associate Professor, 農学部, 助教授 (80038306)
DOIRA Hiroshi Kyushu University, Faculty of Agriculture. Professor, 農学部, 教授 (30038210)
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| Project Period (FY) |
1988 – 1990
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| Keywords | Yellow blood / White blood / Carotenoid binding Protein / CBPL / midgut / CBP-anti-serum / cDNA library / CBP clone |
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| Research Abstract |
1. Biophysical and Biochemical Properties of Carotinoid Binding Protein (H. Fujii and H. Doira)
Carotenoid binding protein like protein (CBPL) was isolated from white blood, CBPL did not contain carotenoid and hence had no color. Molecular weigh of CBPL was 600 kDa the same as that of CPB. Both CBP and CBPL were glycolipoprotein containing carbohydrate and lipid and were made up of two subunits, haevy chain (223 kDs) and light chain (76 kda). In a amino acid compositionof CBP and CBPL, they were characterized by remarkable less methionin, cystein and tyrosine, and much aspartic acid and leucin. By using immuno-western-blotting, CBP was detected on midgut, ovary and egg other than blood. CBP was synthesized in midgut in in vivo and in vitro protein synthesis.
2. Isolation of CBP gene and its structure (Y. Kawaguchi and H. Fujii)
Total RNA containing CBP mRNA was extracted the larval midgut of 3 day of fifth instar by ultra centrifugation using CsF. Poly-A-RNA was isolated from total RNA with oligo dT column. cDNA was synthesized from poly-A RNA as template and cDNA library from poly-A RNA was construct-ed with lambdagll phage. Screening CBP cDNA was carried out using anti-CBP-IgG as probe and one clone was isolated and its molecular size was 1kb. we were working sequence analysis of the clone.
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