@article{oai:iwate-u.repo.nii.ac.jp:00014965, author = {安川, 洋生}, journal = {岩手大学教育学部附属教育実践総合センター研究紀要, The Journal of the Clinical Research Center for Child Development and Educational Practices}, month = {Mar}, note = {データベース解析によりTetX のバリアントをトリの病原菌であるRiemerella anatipestifer から5種見いだした。それらをコードするプラスミドを作成し大腸菌JM109に導入してアッセイしたところ,TetX 発現菌はチゲサイクリン(TGC)に感性を示したが,TetX バリアント発現菌はいずれも耐性を示した。バリアントのうちTetX 5はTetX と20ヶ所のアミノ酸が異なり,この20残基のいずれかが高TGC 耐性に関与していると推察された。そこでTetX5をベースに,これらのアミノ酸を1残基ずつTetX 型に置換した部位特異的変異体を作成しアッセイしたところ,E94,A217,K266,S282のいずれかを置換した変異体はTetX5より有意に耐性度が低下した(p < 0.05, n= ₈ )。次に,TetX をベースに,これら4残基をTetX5型に置換した部位特異的変異体を作成しアッセイしたところTGC 耐性を示し,その耐性度はTetX5による耐性度と有意差が認められなかった。即ち,TetX は4残基を置換することでTetX5と同程度の高TGC 耐性を獲得することが明らかとなった。, Five variants of TetX were found from the avian pathogen Riemerella anatipestifer by database analysis and were named TetX4, TetX5, TetX6, TetX7 and TetX8. Plasmids encoding the genes for TetX variants were constructed and introduced into Escherichia coli JM109 cells, and the transformants were subjected to tigecycline( TGC) sensitivity assays. The disc diffusion assays showed that TetX-expressing cells were sensitive to TGC, but all TetX variant-expressing cells were resistant. Among the variants, TetX5 has the highest homology with TetX. The number of the amino acid residues different between the proteins was 20, speculating that any of these 20 residues was involved in TGC resistance. Twenty site-directed mutants of TetX5 were constructed and subjected to TGC sensitivity assays. The results obtained from the assays suggested that 4 residues in TetX5, E94, A217, K266 and S282, were important for TGC resistance. The site-directed mutant of TetX, TetX113, in which K94, S217, E266 and L282 were changed to E94, A217, K266 and S282, conferred TGC resistance to E.coli cells as did TetX5.}, pages = {105--109}, title = {テトラサイクリン耐性因子TetX とそのバリアントの比較解析}, volume = {19}, year = {2020} }