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首页> 外文期刊>Infection and immunity >Cloning and expression of functional fragment C of tetanus toxin.
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Cloning and expression of functional fragment C of tetanus toxin.

机译：破伤风毒素功能片段C的克隆和表达。

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摘要

A segment of Clostridium tetani DNA corresponding to fragment C of tetanus toxin was amplified by using the polymerase chain reaction. This fragment was cloned into expression vector pTTQ8, under the control of the tac promoter. Expression of this plasmid in Escherichia coli resulted in the production of a protein consisting of 8 amino acids of the vector fused to the C-terminal 460 amino acids of tetanus toxin. This protein (rFragment C) was recognized by an antipeptide antibody specific for fragment C in an enzyme-linked immunosorbent assay and on immunoblots. rFragment C could be purified significantly in one step by immunoaffinity chromatography. Immunization of mice with rFragment C resulted in the production of antibodies that were able to protect the mice against a challenge with tetanus toxin. rFragment C bound to ganglioside GT1b and to neuronal cells in a manner indistinguishable from that of fragment C obtained by papain cleavage of tetanus toxin. For many applications, rFragment C appears to be a suitable alternative to tetanus toxin or toxin-derived fragment C.
机译：通过使用聚合酶链反应扩增破伤风梭菌DNA的对应于破伤风毒素片段C的片段。在tac启动子的控制下，将该片段克隆到表达载体pTTQ8中。该质粒在大肠杆菌中的表达导致产生一种蛋白质，该蛋白质由与破伤风毒素的C端460个氨基酸融合的载体的8个氨基酸组成。在酶联免疫吸附试验和免疫印迹中，该蛋白（rFragment C）被片段C特异的抗肽抗体识别。 rFragment C可以通过免疫亲和色谱一步一步纯化。用rFragment C免疫小鼠会产生能够保护小鼠免受破伤风毒素攻击的抗体。 r片段C与神经节苷脂GT1b和神经元细胞的结合方式与木瓜蛋白酶破伤风毒素的裂解所获得的片段C的方式没有区别。对于许多应用，rFragment C似乎是破伤风毒素或毒素衍生片段C的合适替代品。

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