Development of an efficient transformation and gene expression system for a fastidi-ous oral anaerobe
| Project/Area Number |
23592722
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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 |
Morphological basic dentistry
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| Research Institution | Aichi Gakuin University |
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Principal Investigator |
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| Project Period (FY) |
2011 – 2013
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2013: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2011: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | 嫌気性口腔細菌 / タンネレラ・フォーサイシア / ポルフィロモナス・ジンジバリス / 遺伝子導入 / 発現ベクター / 薬剤耐性カセット / ナチュラルコンピテンス / バイオフィルム / 歯周病原細菌 / 遺伝子破壊 / 自然形質転換 / クロラムフェニコール耐性 / 共用ベクター / 遺伝子導入法 / 薬剤耐性マーカー / エレクトロポレーション / 制限修飾系 |
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| Research Abstract |
A comF-dependent natural competence has been discovered for the first time in an important periodontal pathogen, Tannerella forsythia. Keeping T.forsythia in a biofilm throughout the pr-ocedure allowed efficient DNA uptake and allelic replacement. Based on this finding, a simple method has been established for efficient targeted mutagenesis in T.forsythia. One of the critical conditions affect-ing the efficiency of transformation is the freshness of biofilm, and the recommended subculture duration of input biofilm is 4 days just before transformation. Furthermore, an autonomous chloramphenicol-resista-nt CAT cassette, permF-CAT, has been constructed and successfully applied to T.forsythia. Using permF-CAT, a final version of the common expression vector has been designed which should be functional not only in T.forsythia but also in the other Red Complex anaerobes.
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Report
(4 results)
Research Products
(3 results)
