降钙素原的克隆、表达及鉴定

目的：合成降钙素原的全长基因，表达出降钙素原的融合蛋白并对其鉴定，为PCT的功能研究及单抗制备提供有力的实验基础。 方法 采用多引物人工一步合成基因技术合成降钙素原的全长基因，克隆至原核表达载体PET-20B表达出重组蛋白，将此初步表达的蛋白进行Western blot鉴定。 结果 成功表达降钙素原重组蛋白，Western blot 成功鉴定。 结论 成功表达降钙素原重组蛋白并成功鉴定，为明确降钙素原来源和病理生理作用的研究奠定了基础, 以及为制备其单克隆抗体提供实验材料。

Cloning and expression and identification of procalcitonin

Objective: To synthetize the whole gene of procalcitonin, to express the fusion protein, and to identify the procalcitonin, then to provid the subject for study of the functions and preparation of the monoclonal antibody of procalcitonin. Methods: By using single-step assembly of a gene and entire plasmid from large numbers of oligodeoxyribonucleotides technique, the whole gene of procalcitonin was synthetized, then inserted into the prokaryotic expression vector PET-20B and expressed the recombinant protein. The expression of protein were used to assess with western blot. Results: the expression of the recombinant protein had been succeeded and Western blot analysis showed the recombinant protein of procalcitonin was correct.Conclusion: The success of the expression and identification of the recombinant protein is the investigated foundation of the origin、pathological and physiological functions of procalcitonin. It also provid the subject for preparation of the monoclonal antibody of procalcitonin.