| Item type |
☆紀要論文 / Departmental Bulletin Paper(1) |
| 公開日 |
2022-08-19 |
| タイトル |
|
|
タイトル |
〈論文〉プルーフリーディングポリメラーゼを利用したアレル特異的定量PCR法 |
|
言語 |
ja |
| タイトル |
|
|
タイトル |
<PAPERS and REPORTS> Allele Specific Quantitative PCR Using Proofreading Polymerase |
|
言語 |
en |
| 著者 |
藤田, 亮祐
園川, 舞華
久野, 雅明
藤井, 政幸
|
| 言語 |
|
|
言語 |
jpn |
| キーワード |
|
|
主題 |
Allele Specific Quantitative PCR, Single Base Mutation, KRAS, 2’-OMeRNA Modified Primer |
| 資源タイプ |
|
|
資源タイプ識別子 |
http://purl.org/coar/resource_type/c_6501 |
|
資源タイプ |
departmental bulletin paper |
| ID登録 |
|
|
ID登録 |
10.15100/0000022928 |
|
ID登録タイプ |
JaLC |
| 著者 所属 |
|
|
値 |
近畿大学産業理工学部生物環境化学科 |
| 著者 所属 |
|
|
値 |
近畿大学大学院産業理工学研究科産業理工学専攻 |
| 著者 所属 |
|
|
値 |
近畿大学大学院産業理工学研究科産業理工学専攻 |
| 著者 所属 |
|
|
値 |
近畿大学産業理工学部生物環境化学科; 教授 |
| 著者所属(翻訳) |
|
|
値 |
Department of Biological & Environmental Chemistry, Faculty of Humanity Oriented Science and Engineering, Kindai University |
| 著者所属(翻訳) |
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|
値 |
Biological & Environmental Chemistry Course, Graduate School of Humanity Oriented Science and Engineering, Kindai University |
| 著者所属(翻訳) |
|
|
値 |
Biological & Environmental Chemistry Course, Graduate School of Humanity Oriented Science and Engineering, Kindai University |
| 著者所属(翻訳) |
|
|
値 |
Department of Biological & Environmental Chemistry, Faculty of Humanity Oriented Science and Engineering, Kindai University |
| 版 |
|
|
出版タイプ |
NA |
|
出版タイプResource |
http://purl.org/coar/version/c_be7fb7dd8ff6fe43 |
| 出版者 名前 |
|
|
出版者 |
近畿大学産業理工学部 |
| 書誌情報 |
ja : かやのもり：近畿大学産業理工学部研究報告
en : Reports of Faculty of Humanity-Oriented Science and Engineering, Kindai University
号 33,
p. 49-56,
発行日 2022-06-30
|
| ISSN |
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|
収録物識別子タイプ |
ISSN |
|
収録物識別子 |
13495801 |
| 抄録 |
|
|
内容記述タイプ |
Abstract |
|
内容記述 |
In the present study, quantitative analysis of single nucleotide mutated genes by allele specific real time PCR using chemically modified primers and proofreading DNA polymerases is described. Proofreading DNA polymerase has 3’-exonuclease activity and can correct replication errors and amplify the template DNA with high accuracy if normal phosphate primers are used. In our previous studies, we found that the primers bearing 2’-OMeRNA at the 3’-terminal position completely inhibited polymerase activity of DNA polymerases and that the primers bearing 2’-OMeRNA next to the 3’-terminal position did not inhibit 3’-exonuclease and polymerase activities at all. Therefore, we expected that the mismatched bases in the primers bearing 2’-OMeRNA next to the 3’-terminal position and mismatched bases at the 3’-terminal position would be deleted by 3’-exonuclease activity and the resulting primer bearing 2’-OMeRNA at the 3’-terminal position would stop amplification of the template. As a result, only the primer bearing 2’-OMeRNA next to the 3’-terminal position and a matched base at the 3’-terminal position would be extended. The results were well consistent with our working hypothesis. We found that the primers bearing 2’-OMeRNA next to the 3’-terminal position and a matched base at the 3’-terminal position could amplify the template with similar Ct values to those for normal primers and that the primers bearing 2’-OMeRNA next to the 3’-terminal position and a mismatched base at the 3’-terminal position stopped amplification of the template and no PCR products were obtained. Finally we successfully achieved a novel quantitative PCR method completely discriminating any type of single point mutation at the 35th position of KRAS, KRASG12D, KRASG12A and KRASG12V, using 2’-OMeRNA modified primers and a proofreading polymerase. |
|
言語 |
en |
| フォーマット |
|
|
内容記述タイプ |
Other |
|
内容記述 |
application/pdf |
AA12013209-20220630-0049.pdf (1.7 MB)
