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目的探讨赤鱼工亲环素A基因(dsCypA)的生物学功能。方法构建硫氧还蛋白基因融合表达体系PETTRX-dsCypA,低温诱导表达的融合蛋白dsCypA-TRX,利用金属螯合亲和层析纯化后,经蛋白酶切割和进一步纯化,得到高纯度成熟重组dsCypA,测定其酶活性。结果重组dsCypA具有与人CypA相近的肽基脯氨酸顺反异构酶活性,这在鱼类CypA中是首次报道。结论dsCypA的功能克隆可能部分地解释赤鱼工尾刺中药应用的分子生物学机制,也为从比较生物学的角度,深入开展CypA基因的结构与功能关系研究奠定了基础。
Objective To investigate the biological function of the erythrocyte cyclophilin A gene (dsCypA). Methods The fusion protein dsCypA-TRX was induced by low temperature induction of PETTRX-dsCypA. The purified dsCypA-TRX fusion protein was purified by metal chelate affinity chromatography and purified by protease. Its enzymatic activity. Results Recombinant dsCypA had peptidyl-proline cis-trans isomerase activity similar to human CypA, which was first reported in CypA fish. Conclusion The functional cloning of dsCypA may partly explain the molecular biological mechanism of the application of traditional Chinese medicine Tail of Tail, and lay a foundation for the further study of the relationship between structure and function of CypA gene from the perspective of comparative biology.