1989 Fiscal Year Final Research Report Summary
Basic Studies on Evolution Machines
| Project/Area Number |
62420055
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
生物物性学
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| Research Institution | Saitama University |
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Principal Investigator |
HUSIMI Yuzuru Saitama Univ., Dept. of Environ. Chem., Prof., 工学部, 教授 (80011641)
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| Co-Investigator(Kenkyū-buntansha) |
KIHARA Hiromu Saitama Univ., Dept. of Environ. Chem., Res. Assoc., 工学部, 助手 (90204958)
IIDA Takeaki Saitama Univ., Dept. of Appl. Chem., Prof., 工学部, 教授 (30013171)
NISHIGAKI Koichi Saitama Univ., Dept. of Environ. Chem., Assoc. Prof., 工学部, 助教授 (10107378)
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| Project Period (FY) |
1987 – 1989
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| Keywords | EVOLUTIONARY MOLECULAR ENGINEERING / EVOLUTION REACTOR / CELLSTAT / CONTINUOUS CULTURE OF BACTERIOPHAGE / BASE SEQUENCE SPACE / SELECTIVE VALUE LANDSCAPE / PCR / CELL-SORTER |
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| Research Abstract |
1. Development of an artificial selection-type evolution machine. (1) A cloning system for a specified gene which is inserted into fd DNA has been developed. A specially modified cell-sorter can put a carrier-cell into a well of 1000-well microplate without the free background phage. (2) A quantitative screening method for a cloned mutant gene(lac promoter) has been demonstrated. A fluorogenic substrate RG for beta-galactosidase and a fluorescence microplate-reader were used.
2. Running a natural selection-type evolution machine. (1) Theoretical studies: It is shown that the estimation of selective value landscape on the base sequence space is important. Selective value distribution in mutants population determines whether the selection profile is the two-bodies type or a cascading type. In the case of large mutation rate, the selection is a cooperative phenomenon and the "free selective value" determines the selection. (2) Hybrid phases among various wild-type strains of the phage Ff were made and the selection coefficient of them were measured. An additivity hypothesis of the contribution of each gene to the whole selection coefficient was studied. (3) A fd phage population having a randomly mutagenized major coat protein was made and was partially selected by the celistat culture.
3. Development of DNA handling process. PCR can amplify the specific gene directly from carrier-cells (or probably from a single carrier-cell) in culture medium.
4. Development of a ISFET micro pH senser. By coating the gate surface with polymers we made a difference-type pH senser. We measured time-cources of inorganic pyrophosphatase catalysis in several tens mul solution. An autocatalytic mechanism by Ca ion was elucidated.
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[Publications] Biebricher,C.K., Eigen,M., Gardiner,W., Husimi,Y., Keweloh,H.-C. and Obst,A.: "Modeling Studies of RNA Replication and Viral Infection" Proc. of 2nd Int. Workshop on Modeling of Chem. Reactions, pp.17-38 (Springer, 1987).
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「研究成果報告書概要(欧文)」より
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[Publications] Husimi,Y., Nishigaki,K., Kihara,H., and Kinoshita,Y.: "Selection of Single-Stranded DNA Phage in Cellstat Culture" Proc. of 19th Yamada Conf. on Ordering and Organization in Ionic Solutions, pp.165-175 (World Sci.Pub., Singapore, 1988).
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[Publications] Nishigaki,K., Yamada,T., Watanabe,T., Husimi,Y., and Tanaka,T.: "Bacterial Bioassay of Microgram to Subnanogram Quantities of Glucose" Biotechnol. and Appl.Biochem. 10, 428-434 (1988).
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