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周颖, 薛小燕, 白卉, 马雪, 侯征, 罗晓星. 计算机辅助设计抗MRSA耐药基因mecR1的10-23型脱氧核酶[J]. 中国药科大学学报, 2010, 41(4): 380-384.
引用本文: 周颖, 薛小燕, 白卉, 马雪, 侯征, 罗晓星. 计算机辅助设计抗MRSA耐药基因mecR1的10-23型脱氧核酶[J]. 中国药科大学学报, 2010, 41(4): 380-384.
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Computer-aided design of the 10-23 deoxyribozyme targeting resistance gene mecR1[J]. Journal of China Pharmaceutical University, 2010, 41(4): 380-384.
Citation: Computer-aided design of the 10-23 deoxyribozyme targeting resistance gene mecR1[J]. Journal of China Pharmaceutical University, 2010, 41(4): 380-384.
shu

计算机辅助设计抗MRSA耐药基因mecR1的10-23型脱氧核酶

Computer-aided design of the 10-23 deoxyribozyme targeting resistance gene mecR1

    摘要
  • 摘要: 应用Primer Premier 5.0和RNA structure 4.6两种软件设计和筛选有效抗mecR1的10-23型脱氧核酶（DRz），采用电转化的方法将其导入细菌体内，实时PCR的方法定量检测其对mecR1转录的影响，并通过平板克隆形成实验观察脱氧核酶抑制细菌生长的情况。结果表明，计算机辅助设计合成的5条抗MRSA耐药调控基因mecR1的10-23型脱氧核酶，它们能不同程度的降低mecR1的转录水平，有效抑制临床耐药菌MRSA080309的生长，其中DRz6抑制效果最显著。说明联合应用这两种计算机软件设计抗mecR1的10-23型脱氧核酶，是一种经济实用而有效的方法，能够大大缩短有效反义药物的筛选时间。

     

    Abstract: The purpose of this study was to exploit the potential of Primer Premier 5.0 and RNA structure 4.6 in the design and selection of effective 10-23 deoxyribozyme (DRz) targeting resistance gene mecR1.Five designed and synthesized anti-mecR1 10-23 DRz sequences were introduced into the MRSA strain by electro transformation in vivo.Transcription of mecR1 was analyzed by real-time quantitative PCR.The inhibitory effects of DRzs on the bacterial growth were evaluated based on the plate cloning formation.It was found that the five anti-mecR1 10-23 DRz sequences significantly lowered the transcription of mecR1 and inhibited the growth of clinical drug-resistant MRSA080309，with DRz6 having the most significant inhibitory effect.Therefore，the combination of the two computer softwares is an economical，practical and effective approach in the design of anti-mecR1 DRz，and could greatly reduce the screening time of antisense drugs.

     

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