1991 Fiscal Year Final Research Report Summary
Developmetn of a method for cloning of cDNAs by phenotypic complementation of mammalian or fission yeast cells
| Project/Area Number |
01890009
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
広領域
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| Research Institution | Osaka University |
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Principal Investigator |
OKAYAMA Hiroto Osaka University, Research Institute for Microbial Diseases, Professor, 微生物病研究所, 教授 (40111950)
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| Co-Investigator(Kenkyū-buntansha) |
ONO Yasuko Osaka University, Research Institute for Microbial Diseases, Technical Instructo, 微生物病研究所, 教務職員(文部技官) (70194602)
NAGATA Akihisa Osaka University, Research Institute for Microbial Diseases, Instructor, 微生物病研究所, 助手 (50155933)
NOJIMA Hiroshi Osaka University, Research Institute for Microbial Diseases, Associate Professor, 微生物病研究所, 助教授 (30156195)
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| Project Period (FY) |
1989 – 1991
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| Keywords | cDNA / Expression Cloning / Schizosacchromyces Pombe / Heterologous Genetic Complementation / Cell Cycle / cdc25 / Cdc2 / Weel |
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| Research Abstract |
We have developed a method that permits cloning of CDNAs based on. phenotypic complementation of mammalian or fission yeast cells. This method is composed of 1)a high-efficiency full-length CDNA cloning method ; 2)the pcD2 expression vector ; 3)a high-frequency mammalian cell transfection method ; 4)a high-efficiency yeast transfection method ; 5)highly-stable yeast vectors for transducing CDNA libraries into fission yeast.
Using this method, we have isolated two distinct human and rat homologs of the cdc25^+ gene, a fission yeast mitotic inducer, and human and rat homologs of the weel^+ gene, a fission yeast mitotic inhibitor. In addition we have isolated several human genes that suppress temperal'ure-sensitive cdc2 mutants or/and a temperature-sensitive sexual differentiation-proficient patl mutant. Some of them encodes a protein with RNA-bindng domains and some others a protein which. might associate with cdc2 kinase.
This CDNA cloning system will be useful for the islation of a wide variety of mammalian genes.
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