| Project/Area Number |
10306013
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Fisheries chemistry
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
NISHITA Kiyoyoshi Hokkaido Univ., Grad. School of Fisheries Sciences, Professor, 大学院・水産科学研究科, 教授 (20001620)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Hiroyuki Hokkaido Univ., Grad. School of Fisheries Sciences, Instructor, 大学院・水産科学研究科, 助手 (90241372)
OJIMA Takao Hokkaido Univ., Grad. School of Fisheries Sciences, Associate Professor, 大学院・水産科学研究科, 助教授 (30160865)
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| Project Period (FY) |
1998 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥8,380,000 (Direct Cost: ¥7,900,000、Indirect Cost: ¥480,000)
Fiscal Year 2001: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2000: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1999: ¥3,600,000 (Direct Cost: ¥3,600,000)
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| Keywords | myosin / actin / amino acid sequenc / cDNA cloning / fish / salmon / arabesque greenling / walleye pollack / ミオシン重鎖 / サバ / 心筋タイプ / 骨格筋タイプ / 塩基配列 / 筋タンパク質 / 多獲魚 |
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| Research Abstract |
The cDNA libraries were constructed from dorsal muscle of four species of fishes. In case of salmon, the libraries were also constructed from dark and cardiac muscles. Then, the cDNAs for the myosin heavy chains and actins were cloned and amplified by PCR. Nucleotide sequences were determined by thermal cycle sequencing. Results obtained were as follows;
1. Two species of myosin heavy chain cDNA clones were found in the salmon ordinary muscle. The nucleotide sequences of the both clones were determined and deduced the complete amino acid sequences of 1,937 residues. The sequences show low homology as 71 % and dispersed amino acid replacements. 2. The myosin cDNA clone was prepared from salmon dark muscle and cardiac muscle. From their nucleotide sequences, amino acid sequences of 246 residues were deduced According to the sequence comparison and database search, one of the clones found in the salmon ordinary muscle was concluded to be identical with the dark muscle myosin clone. 3. From the cDNA library of the ordinary muscle of arabesque greenling, two species of cDNA clones encoding ordinary muscle myosin and dark muscle myosin were obtained. Thereby, complete amino acid sequence of 1931 residues of the "dark muscle type" myosin was determined. The sequence of 832 residues of the "dark muscle type" was also determined. 4. As for the mackerel, partial amino acid sequences of 245 residues were determined of the both ordinary and dark muscle type myosins. 5. Three species of actin cDNA clones of salmon, walleye pollack, and arabesque greenling were obtained and determined the nucleotide sequences deducing the complete amino acid sequences of 375 residues. 6. The sequence comparison and discussion on the fish myosins and actins were made from various viewpoints. Two manuscripts were made and submitted.
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