体外转录 CRISPR 相关 RNA 降低铜绿假单胞菌耐药性的研究

摘要

目的：利用铜绿相假单胞菌 CRISPR 相关 RNA(crRNA)干扰其耐药基因的表达，以降低泛耐药菌株的耐药性。方法：体外转录获得靶向铜绿假单胞菌耐药相关基因 rhlI 的 crRNA，热激法将 crRNA 转化进入铜绿假单胞菌，纸片扩散法测定细菌转化前后的抑菌圈直径，采用非参数估计的配对秩和检验对药敏结果进行统计学分析。结果：成功转录出针对 rhlI 基因的 crRNAs，且铜绿假单胞菌转化后抑菌圈的直径要显著大于转化前的直径。结论：体外转录的 crRNAs 可以降低铜绿假单胞菌泛耐药菌株的耐药性。%ObjectiveUsing CRISPR associated RNA (crRNA) of Pseudomonas aeruginosa to interfere with the expression of drug resistant gene, in order to reduce antibiotic resistance of drug-resistance Pseudomonas aeruginosa strains. Methods Transcribing crRNA targeting rhlI gene which contribute the drug resistance of Pseudomonas aeruginosa in vitro, transforming crRNA into Pseudomonas aeruginosa by heat shock, and taking the K-B disc diffusion test and then comparing the diameter of inhibition zone by Wilcoxon signed-rank test of non-parametric estimation statistically.Results The crRNAs were transcribed, and the strains which were transformed crRNAs statistically show a bigger inhibition zone than those not.Conclusion The crR-NAs transcribed in vitro could reduce antibiotic resistance of drug-resistance Pseudomonas aeruginosa strains.