降钙素原的克隆、表达及多抗制备

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制备多抗，为降钙素原（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pGEX-4T-1表达出重组蛋白，将初步纯化的蛋白免疫小鼠制备多抗并进行Western blot鉴定。结果成功表达降钙素原重组蛋白并制备其多抗，Western blot证实所制备的PCT多抗可与所获得的融合蛋白和病人血清发生特异性反应。结论成功表达降钙素原重组蛋白并制备其多抗，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,Expression and Production of Polyclonal Antibody to Procalcitonin

Objective To construct a recombinant expression plasmid containing the whole coding sequence of procalcitonin, to express the fusion protein, and to produce the polyclonal antibody of procalcitonin. Methods The whole coding region of procalcitonin gene was chemically synthesized, assembled, and then inserted into the prokaryotic expression vector pGEX-4T-1. The recombinant protein was expressed in E.coll. The purified protein was used as immunogen to prepare the polyclonal antibody and as target protein for western blot. Results Recombinant procalcitonin was successfully expressed. Antibody to this recombinant protein was also produced. Western blotting analysis showed that antibody to this recombinant protein was specific for procalcitonin and the patient sera. Conclusion Recombinant procalcitonin and antibody to this protein was successfully produced. With this recombinant protein, the research on the pathological and physiological functions of procalcitonin would be facilitated.