中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2009年
11期
1009-1013
,共5页
陈茶%黄彬%薛新娜%林冬玲%罗强%蓝锴%鄂顺梅
陳茶%黃彬%薛新娜%林鼕玲%囉彊%藍鍇%鄂順梅
진다%황빈%설신나%림동령%라강%람개%악순매
10-23脱氧核酶%铜绿假单胞菌%VIM2%亚胺培南%基因治疗
10-23脫氧覈酶%銅綠假單胞菌%VIM2%亞胺培南%基因治療
10-23탈양핵매%동록가단포균%VIM2%아알배남%기인치료
10-23 deoxyribozyme%Pseudomonas aeruginosa%VIM2%Imipenem%Gene therapy
目的 探讨10-23脱氧核酶(DRz)抑制铜绿假单胞菌耐药基因VIM2的表达及其在逆转铜绿假单胞菌耐药中的作用.方法 根据计算机模拟的 VIM2 mRNA 的二级结构设计合成 5 条VIM2特异的10-23DRz(DRz1～DRz5),在无细胞反应体系中鉴定10-23DRz对VIM2 mRNA的切割活性后,将其导入铜绿假单胞菌,利用E-test法检测亚胺培南对处理前后铜绿假单胞菌的MIC值.结果 DRz3、DRz4和DRz5在无细胞反应体系中可对VIM2 mRNA进行有效切割,且其活性具有高度特异性.与对照相比,1O-23DRz可以降低亚胺培南对铜绿假单胞菌的MIC值.结论 实验中所设计的10-23DRz能在细胞外高效特异性的切割VIM2 mRNA;在细胞内能协同业胺培南抑制铜绿假单胞菌耐药.
目的 探討10-23脫氧覈酶(DRz)抑製銅綠假單胞菌耐藥基因VIM2的錶達及其在逆轉銅綠假單胞菌耐藥中的作用.方法 根據計算機模擬的 VIM2 mRNA 的二級結構設計閤成 5 條VIM2特異的10-23DRz(DRz1～DRz5),在無細胞反應體繫中鑒定10-23DRz對VIM2 mRNA的切割活性後,將其導入銅綠假單胞菌,利用E-test法檢測亞胺培南對處理前後銅綠假單胞菌的MIC值.結果 DRz3、DRz4和DRz5在無細胞反應體繫中可對VIM2 mRNA進行有效切割,且其活性具有高度特異性.與對照相比,1O-23DRz可以降低亞胺培南對銅綠假單胞菌的MIC值.結論 實驗中所設計的10-23DRz能在細胞外高效特異性的切割VIM2 mRNA;在細胞內能協同業胺培南抑製銅綠假單胞菌耐藥.
목적 탐토10-23탈양핵매(DRz)억제동록가단포균내약기인VIM2적표체급기재역전동록가단포균내약중적작용.방법 근거계산궤모의적 VIM2 mRNA 적이급결구설계합성 5 조VIM2특이적10-23DRz(DRz1～DRz5),재무세포반응체계중감정10-23DRz대VIM2 mRNA적절할활성후,장기도입동록가단포균,이용E-test법검측아알배남대처리전후동록가단포균적MIC치.결과 DRz3、DRz4화DRz5재무세포반응체계중가대VIM2 mRNA진행유효절할,차기활성구유고도특이성.여대조상비,1O-23DRz가이강저아알배남대동록가단포균적MIC치.결론 실험중소설계적10-23DRz능재세포외고효특이성적절할VIM2 mRNA;재세포내능협동업알배남억제동록가단포균내약.
Objective To investigate the inhibitory effects of 10-23 deoxyribozyme (DRz) on expression of VIM2 and the effect against tolerance of Pseudomonas aeruginosa. Methods Five 10-23DRzs targeting VIM2 gene( DRzl-DRz5) were designed according to the predicted secondary structure of VIM2 mRNA. Their cleavage activity and specificity were identified in cell free conditions. 10-23DRz were introduced into Pseudomonas aeruginosa producing β-lactamas by electroporation procedure. Following electroporation, MICs of imipenem against Pseudomonas aeruginosa were detected by E-test method. Results Three of the five designed 10-23DRz (DRz3-DRz5) could cleavage VIM2 mRNA efficiently and specifically. MIC values aginst Pseudomonas aeruginosa were lower than that in control DNA electroporated. Conclusion 10-23DRz can cleave VIM2 mRNA with effectiveness and specificity, and exhibit synergic effect with imipenem on Pseudomonas aeruginosa, which will probably play an important role in the gene therapy of tolerance.
