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目的:研究互补于MRSA的耐药基因mecR1mRNA的PSODNs6088对MRSA耐药性的影响.方法:利用电穿孔的方法将PSODNs6088导入MRSA内,平板克隆形成实验计数菌落数(CFU);微量法测定细菌的生长曲线;RTPCR法检测目的基因mecR1和mecA的表达变化.结果:PSODNs6088能明显抑制MRSA的生长,PSODNs6088各剂量组MRSA的CFU与空白对照组比较明显减少(P<0.05),且具有剂量依赖性,而随机对照组的菌落数却无明显减少;PSODNs6088能显著抑制MRSAmecR1和mecA的表达.结论:反义硫代寡核苷酸能特异的与mecR1mRNA结合并阻断其表达,从而阻断了MRSA耐药基因表达信号通路,减少MRSA的菌落数,抑制其生长,提示反义寡核苷酸能有效的逆转MRSA对β内酰胺类抗生素的耐药性.利用反义技术阻断细菌的耐药信号通路可能成为解决细菌耐药问题的一种新策略.
Objective: To study the effect of PSODNs6088 on the MRSA resistance of drug resistance gene mecR1 mRNA complementary to MRSA.Methods: PSODNs6088 was introduced into MRSA by electroporation, and the number of colonies was counted to form experimental count colonies (CFU) The growth curve was detected by RT-PCR and the expression of mecR1 and mecA were detected by RTPCR method.Results: PSODNs6088 could significantly inhibit the growth of MRSA.Compared with blank control group, the CFU of MRSA in each dose group of PSODNs6088 significantly decreased (P <0.05) , While there was no significant reduction in the number of colonies in the randomized control group.PSODNs6088 could significantly inhibit the expression of MRSAmecR1 and mecA.Conclusion: Antisense oligonucleotides can specifically bind to mecR1 mRNA and block its expression, thus blocking MRSA The results indicated that antisense oligodeoxynucleotides can effectively reverse the resistance of MRSA to β-lactam antibiotics.Using antisense technology to block the resistance of bacteria The signaling pathway may be a new strategy to solve the problem of bacterial resistance.