1995 Fiscal Year Final Research Report Summary
Studies of structure and physiological function of chymotrypsin inhibitors in hemolymph of Bombyx mori
| Project/Area Number |
05454066
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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・昆虫利用学
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| Research Institution | Kyushu University |
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Principal Investigator |
FUJII Hiroshi Kyushu University, Agriculture, Associate Professor, 農学部, 助教授 (10038268)
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| Co-Investigator(Kenkyū-buntansha) |
ASO Yoichi Kyushu University, Agriculture, Associate Professor, 農学部, 助教授 (10117054)
KAWAGUCHI Yutaka Kyushu University, Agriculture, Associate professor., 農学部, 助教授 (80038306)
KOGA Katsumi Kyushu University, Agriculture, professor, 農学部, 教授 (40038261)
DOIRA Hiroshi Kyushu University, Agriculture, professor, 農学部, 教授 (30038210)
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| Project Period (FY) |
1993 – 1995
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| Keywords | Bombyx mori / chymotrypsin inhibitor (CI) / CI-1 amino acid sequence / CI-lb amino acid sequence / CI-8 cDNA sequence / fat body cDNA library |
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| Research Abstract |
(1) Determination of amino acid sequence of chymotrypsin inhibitor (Fujii and Aso) :
Basic chymotrypsin inhibitors, CI-1 and CI-b1 controlled by Ict-A complex gene were isolated from larval hemolymph of the silkworm, Bombyx mori according to the purification method established by us. Isolated CI-1 consisted of 62 amino acid and its molecular weight was calculated 7,065 on the basis of the amino acid sequence. Number of amino acid of CI-b1 was equal to the CI-1. (also consisted of 62 amino acids as well as CI-1). and a molecular weight of 7,065. Both CIs were sequenced amino acid. The amino acid sequences of CI-1 and CI-b1 were followed ; .
CI-1 DKPTTEPICEQAFGNSGPCFAYIKLYSYNQKTKKCEEFIYGGCQGN
CI-b1 -----K-----------K---
DNRFITLAECEQKCIK
----D-------
Glu at the 6th amino acid of CI-1 was changed into Lys at CI-b1, Gln at the 44th into Lys, and Ile at the 51st into Asp.
(2) Analysis of structure of chymotrypsin inhibitor, CI-8 gene (Fujii and Doira) :
CI-8 cDNA was amplified by PCR with the fat body cDNA library using the synthesized oligonucleotides as probe based on both sequences of N-terminal and C-terminal amino acid sequences. CI-8 cDNA was cloned to pUC9 plasmid. and was sequenced nucleotide. It consisted of 1,185bp and the deduced amino acid sequence consisted of 395a.a. CI-8 cDNA contained the signal peptide consisted of 16 amino acids being rich with lyophilic amino acid such as Leu and Val.
(3) Analysis of physiology of chymotrypsin inhibitors (Kawaguchi and Koga) :
The radioisotope labeled CI-8 was injected into the 5th instar larvae. Abundant labeled CI-8 was detected in the fat body resident along epidermis and did not in the fat body near the midgut.
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