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微生物学通报

抗菌肽BuforinⅡ衍生物抑制大肠杆菌大分子合成的研究
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国家自然科学基金项目(No. 31172214); 中央高校基本科研业务基金项目(No. 11NZYQN30); 国家十二五支撑计划项目(No. 2012BAD33B05)


Two analogues of the antimicrobial peptide buforinⅡ inhibit bacterial macromolecular synthesis in Escherichia coli
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    摘要:

    【目的】研究抗菌肽BuforinⅡ的衍生物BF2-A/B作用大肠杆菌后对胞内生物大分子合成的影响。【方法】紫外分光光度法检测抗菌肽对细胞DNA、RNA合成能力的影响, 考马斯亮蓝法检测抗菌肽对细胞总蛋白合成能力的影响, 分别用邻硝基苯-β-D-吡喃半乳糖苷法和对硝基苯磷酸二钠法检测抗菌肽对β-半乳糖苷酶及碱性磷酸酶表达活性的影响。【结果】BF2-A/B不优先抑制DNA合成, 而是优先抑制RNA和蛋白的合成。在相同浓度下, BF2-B抑制RNA合成的能力比BF2-A强, BF2-A抑制蛋白合成的能力比BF2-B强。胞内酶β-半乳糖苷酶和碱性磷酸酶的表达活性都在下降。【结论】BF2-A/B结合胞内核酸后, 没有首先影响DNA的复制功能, 而是优先抑制基因转录功能, 主要在转译水平上抑制蛋白的合成。

    Abstract:

    [Objective] In this paper, the influence to bacteria intracellular macromolecular biosynthesis after antimicrobial peptides BF2-A and BF2-B, two analogues of buforin Ⅱ, interacted with E. coli had been researched. [Methods] The influence of peptides on the synthesis of bacterial DNA and RNA had been determined by ultraviolet spectrophotometry. The influence of peptides on the synthesis of bacterial total protein had been determined by Coomassie brilliant blue method. Inhibition of β-galactosidase and alkaline phosphatase activities of E. coli cells by antibacterial peptides had been measured by ONPG and PNPP, respectively. [Results] Peptides didn’t primarily influence the synthesis of DNA and there was a lag phase. Thus the preferential inhibition of biosynthesis of RNA and protein appeared to be the primary target of BF2-A/B and the effect of the peptides on DNA-synthesis appeared to be secondary. On the same concentration, BF2-B caused a more marked inhibition of RNA-synthesis than BF2-A. Interestingly, BF2-A induced a more remarkable inhibition of protein synthesis than BF2-B. The results of activity measuring of two endocellular enzymes, β-galactosidase and alkaline phosphatase, further proved that the biosynthesis of protein was inhibited. [Conclusion] Both peptides binding to nucleic acid didn’t primarily inhibit the replication function of DNA, but primarily inhibit the transcription function of gene. Moreover, the preferential inhibition of protein-synthesis mostly embodied at the translation level.

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郝刚,施用晖,马丹雅,乐国伟. 抗菌肽BuforinⅡ衍生物抑制大肠杆菌大分子合成的研究[J]. 微生物学通报, 2013, 40(11): 2057-2065

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  • 在线发布日期: 2013-11-04
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