Cloning of characean myosin
| Project/Area Number |
08640819
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
植物生理
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| Research Institution | Chiba University |
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Principal Investigator |
YAMAMOTO Keiichi Chiba University Biology Professor, 理学部, 教授 (70053361)
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| Co-Investigator(Kenkyū-buntansha) |
MIMURA Tetsurou Hitotsubashi Univ.Business Professor, 教授 (20174120)
YAJIMA Hirohiko Chiba University Biology Res.Assist., 理学部, 助手 (30261895)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1996: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | charophyta / myosin / cytoplasmic streaming / molecular cloning / ミオンン / シャジクモ |
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| Research Abstract |
It is well known that characean cytoplasmic streaming is extremely is fast. Although this streaming is known to be generated by myosin-actin system like muscle contraction, shape and characteristics of characean myosin are not understood well. We, therefore, started to clone the cDNA encoding this myosin hoping that sequence information reveals the secret of the world fastest myosin. We, at first, purified characean myosin biochemically and raised antibody againstit. Then, we screened the cDNA library of Chara corallina. We obtained several cDNA clones which cover whole coding region of the heavy chain of characean myosin. Coding region contained 6471 bases corresponding 2157 amino acid residues. Calculated molecular weight was 245532 and this value fitted quite well with that determined by SDS polyacrylamide gel electrophoresis of biochemically purified characean myosin. Secondary structure prediction program indicated that this myosin has, from N-terminus, globular structure with ATPase and actin binding sites, IQ motifs which are light chain binding sites, coiled coil region which favors dimer formation, and globular tail piece. Over all structure indicated agreed very well with the electron microscopic image of this myosin obtained previously by us. We expressed coiled coil region and tail piece in E.coli and raised antibody against these peptides. These antibody recognized biochemically purified characean myosin. These results suggested definitely that the cDNA we cloned represents that of characean myosin.
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Report
(3 results)
Research Products
(11 results)
