Determination of recognition sequence of DNA methyltransferase M. Ape from Hyperthermophilic archaea Aeropyrum pernix
| Autor: | Hayashi, Mao, Sugawara, Keisuke, Iida, Yasuhiro |
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| Jazyk: | japonština |
| Rok vydání: | 2021 |
| Předmět: | |
| Zdroj: | 神奈川工科大学研究報告.B,理工学編. 45:49-55 |
| ISSN: | 2188-2878 |
| Popis: | application/pdf DNA methylation, one of the epigenetic modifications, is mainly reported N6-methyladenine(m6A), N5-methylcytosine(m5C) and N4-methylcytosine(m4C) and there exist in all of the domain of eukaryote, bacteria and archaea. The significant function of DNA methylation is gene regulation, control of cell cycle and Restriction-Modification system (R-M system) of bacteria. The Type II of R-M system is constituted by restriction enzyme and DNA methyltransferase and there recognize same sequence. In the R-M system of hyperthermophilic archaea Aeropyrum pernix, restriction enzyme already has been sale from New England Biolabs Inc., however, the DNA methyltransferase of the pair of restriction enzyme is only demonstrated by homology analysis on genome. In this report, we clarified that the properties of DNA methyltransferase M. Ape of R-M system Type II from Aeropyrum pernix. The ORF sequence of M. Ape was introduced into pCold I vector and expression in E. coli JM109 strain. And then, it was purified by His-tag. The recognition sequence of M. Ape was decided by methylation activity and bisulfite sequence. As the result, it is clarified that M. Ape is cytosine-specific methyltransferase and it modify the second cytosine in GCTGC sequence. And also, it was suggested that the optimum temperature of M. Ape was 60 degrees Celusius. |
| Databáze: | OpenAIRE |
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