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目的对A型肉毒毒素重链C端多肽原始基因序列进行密码子优化后转入大肠杆菌系统进行原核表达并纯化,研究其在多抗血清中的抗原性。方法使用http://www.jcat.de/数据库等生物信息学技术对A型肉毒毒素重链C端多肽原始基因序列进行密码子优化后装入原核表达载体p ET-22b(+)中,转入大肠杆菌工程菌株BL21(DE3)表达并纯化,通过动物实验研究其在多抗血清中的抗原性。结果成功表达并纯化了A型肉毒毒素重链C端多肽,并对其抗原性进行验证。结论 A型肉毒毒素重链C端多肽作为抗肉毒毒素的二代亚单位疫苗组分以及肉毒类毒素的替代品,在制备以及激起中和抗体的方面具有明显的优势。
OBJECTIVE: To optimize the codon usage of the C-terminal polypeptide of botulinum toxin type A botulinum toxin and transfer into Escherichia coli system for prokaryotic expression and purification to study its antigenicity in multi-antiserum. Methods Using the bioinformatics technology such as http://www.jcat.de/ database, the original gene sequence of C-terminal polypeptide of botulinum toxin type A was codon optimized and inserted into prokaryotic expression vector p ET-22b (+) , Transformed into E. coli strain BL21 (DE3), expressed and purified, and its antigenicity in multiple antisera was studied through animal experiments. Results The C-terminal polypeptide of botulinum toxin type A was successfully expressed and purified, and its antigenicity was verified. Conclusions As a second generation subunit vaccine against botulinum toxin and as a substitute for botulinum toxin, the C-terminal polypeptide of botulinum toxin type A has obvious advantages in the preparation and elicitation of neutralizing antibodies.