FUNCTIONAL ANALYSIS OF VISUAL PIGMENTS BY CASSETTE CHIMERA METHOD
| Project/Area Number |
05454633
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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 |
Biophysics
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| Research Institution | Osaka University |
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Principal Investigator |
TOKUNAGA Fumio OSAKA UNIVERSITY,EARTH AND SPACE SCIENCE,PROFESSOR, 理学部, 教授 (80025452)
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| Co-Investigator(Kenkyū-buntansha) |
HISATOMI Osamu OSAKA UNIVERSITY.BIOLOGY RESEARCH ASSOCIATE, 理学部, 助手 (60231544)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥7,100,000 (Direct Cost: ¥7,100,000)
Fiscal Year 1994: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1993: ¥5,900,000 (Direct Cost: ¥5,900,000)
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| Keywords | VISUAL PIGMENT / VISION / RHODOPSIN / COLOR VISION / CHIMERA PROTEIN / EXPRESSION IN CULTURE CELLS / MODIFICATION AFTER TRANSLATION / COLOR / カセット・キメラ法 / キメラ / アミノ酸配列 / 蛋白質工学 / DNA組換え |
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| Research Abstract |
In order to understand the relationship between amino acid sequences of visual pigments and their functions, we cloned cDNAs of various visual pigments, and constructed chimera visual pigments with the cloned cDNAs based on the cDNA of bovine rhodopsin and analyzed the functions of the produced pigments.
1.We cloned cDNAs of frog rhodopsin and green rod visual pigment, and killifish rhodopsin and four cone visual pigments. 2.The frog and bovine rhodopsin and chicken red-sensitive pigments expressed in culture cells showed the same properties as the native ones. Goldfish UV-pigments was not detected in the culture cells. Killifish blue-sensitive pigment expressed in culture cells showed the absorption maximum aroud 450 nm. Bovine rhodopsin N20Q showed the absorption maximum at 500 nm and was glycosylated with different chain lengths. N15Q and E113Y was not detected in the culture cells. Bovine rhodopsin C322/332S had the same properties as the native one on the sbsorption maximum, photos
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ensitivity and resistance against hydroxyl amine, but the rate of the formation metarhodopsin was faster than the native one. 3.Chimera visual pigments were constructed by substituting other visual pigment polypeptides between 258 and 295 of bovine rhodopsin polypeptide. In the cases of goldfish uv-pigment, chicken red-, green-and blue-sensitive pigments, the expressed peptides were not detected in the culture cells. Chimera rhodopsin between bovine and lamprey rhodopsins showed the same properties as the native bovine rhodopsin. In this study we succeeded the expression of visual pigments in the culture cells and could analyze some properties of the expressed visual pigments, but did not detected the pigments in the some cases. We have to improve the expression systems in the near future. We did not observe large shift of the absorption maxima and sensitivity change against hydroxyl amine on chimera visual pigments. These properties may be supported by cooperative interactions of many amino acid residues. Less
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Report
(3 results)
Research Products
(27 results)
