Development of production method of large size of combinatorial libraries by mating of Sccharomyces cerevisiae cells and screening
| Project/Area Number |
12650787
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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 |
生物・生体工学
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| Research Institution | KOBE UNIVERSITY |
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Principal Investigator |
KATOH Shigeo Kobe University, Graduate School of Science and Technology, Professor, 自然科学研究科, 教授 (20026272)
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| Co-Investigator(Kenkyū-buntansha) |
SHIOMI Naofumi Kobe Colledge, Dept. Human Science, Associate Professor, 人間科学部, 助教授 (20299077)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Combinatorial library / Antibody / Production by secretion / Yeast cell / Mating / Screening / メティング |
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| Research Abstract |
Recent years a variety of bioactive materials are widely used in many biological fields. To make sure obtaining bioactive materials with desired characteristics by screening, it is necessary to make huge libraries of these biomaterials. Numbers of species in a library, however, are limited depending on construction methods of libraries.
In this work we try to devise a new method for production of a larger size of combinatorial libraries by use of mating characteristics of yeast cells. There are two kinds of mating types in S. cerevisiae, a-type and α-type, and they can make a diploid, which carries both the genes originated from a-type and α-type. For example, by transforming libraries of genes of heavy and light chains to a- and α-types respectively, it should be possible to obtain a larger size of libraries of antibodies by mating them. Further, since antibodies produced can be designed to be secreted from transformants, straines for production of antibodies may be easily screened.
To confirm our method, the expression vectors containing heavy chain gene (μ) and light chain genes (κ) of human antibody against exotoxin were transformed into a-type and α-type strains of S. cerevisiae, respectively and these transformants were mated by mixing the cultured cells of both types. Secretion of completely assembled antibody was confirmed. It was shown that a library can be doubled by this method. Further, the effects of media, cell concentration and the ratio of a-type cell to α-type cell on the mating efficiency were studied.
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Report
(3 results)
Research Products
(3 results)
