Divisions of Microbiology, Center for Food Safety and Applied Nutrition, Food and Drug Administration, College Park, MD 20740, USA.
Protein J. 2013 Apr;32(4):288-96. doi: 10.1007/s10930-013-9486-1.
Botulinum neurotoxins are produced as a toxin complex (TC) which consists of neurotoxin (NT) and neurotoxin associated proteins. The characterization of NT in its native state is an essential step for developing diagnostics and therapeutic countermeasures against botulism. The presence of NT genes was validated by PCR amplification of toxin specific fragments from genomic DNA of Clostridium botulinum strain PS-5 which indicated the presence of both serotype A and B genes on PS-5 genome. Further, TC was purified and characterized by Western blotting, Digoxin-enzyme linked immunosorbent assay, endopeptidase activity assay, and Liquid chromatography-Mass spectrometry. The data showed the presence of serotype A specific neurotoxin. Based on the analysis of neurotoxin genes and characterization of TC, PS-5 strain appears as a serotype A (B) strain of C. botulinum which produces only serotype A specific TC in the cell culture medium.
肉毒梭菌神经毒素以毒素复合物（TC）的形式产生，该复合物由神经毒素（NT）和神经毒素相关蛋白组成。在天然状态下对 NT 的特性进行表征是开发针对肉毒中毒的诊断和治疗对策的重要步骤。通过从肉毒梭菌 PS-5 菌株的基因组 DNA 中扩增毒素特异性片段的 PCR 扩增验证了 NT 基因的存在，这表明 PS-5 基因组上存在 A 型和 B 型两种基因。此外，通过 Western blot、Digoxin-酶联免疫吸附试验、内肽酶活性测定和液相色谱-质谱对 TC 进行了纯化和表征。结果表明存在 A 型特异性神经毒素。基于神经毒素基因的分析和 TC 的表征，PS-5 菌株似乎是 C. botulinum 的 A 型（B 型）菌株，仅在细胞培养基中产生 A 型特异性 TC。
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