2017 Fiscal Year Final Research Report
Elucidation of the mecahsim of thermostabilization of thermostable reverse transcriptase
| Project/Area Number |
15K07388
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied biochemistry
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| Research Institution | Kyoto University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
兒島 憲二 京都大学, 農学研究科, 助教 (40542759)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Keywords | リボヌクレアーゼH / RNase H / RNase H2 / 逆転写酵素 / GPMA method |
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| Outline of Final Research Achievements |
Ribonuclease H (RNase H) is an enzyme that specifically degrades RNA from RNA/DNA hybrids. RNase H has been classified into two major families, type 1 and type 2. Retroviral reverse transcriptase (RT) has DNA polymerase and RNase H activities. In this study, we aimed to elucidate the RNase H and engaged in the following four subjects. (i) The RNase H domain of MMLV RT was prepared and characterized. (ii) The effects of neutral salts and pH on the activity and stability of human RNase H2 were analyzed. (iii) The GPMA method using mouse NIH3T3 cell line instead of E. coli was established. (iv) The RNase H2 knock-out NIH3T3 cell line was established and characterized.
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| Free Research Field |
酵素化学
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