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目的成功的表达鼠的PrP,制备特异性的兔抗PrP多克隆抗体。方法以BALB/c小鼠肝脏基因组DNA作为模板运用PCR技术成功扩增了PrP23-231,将其克隆到pRSET原核表达载体中,构建重组表达载体pRSET PrP23-231。将该重组载体进行无细胞表达,然后对成功表达的PrP进行纯化,然后用纯化的蛋白免疫新西兰大耳白兔制备其多克隆抗体,并进行Western-blot鉴定。结果通过Western-blot鉴定,有约30kD的目的条带,证明蛋白成功表达,并进行其多克隆抗体的制备并进行Western-blot鉴定,有约30kD的目的条带,证明成功制备了PrP的多克隆抗体。结论成功表达了PrP23-231,并且成功制备了其多克隆抗体,为进一步研究PrP的功能奠定了基础。
OBJECTIVE: To successfully express murine PrP, a specific rabbit anti-PrP polyclonal antibody was prepared. Methods PrP23-231 was successfully amplified by PCR from the liver genomic DNA of BALB / c mice and cloned into prokaryotic expression vector pRSET to construct the recombinant expression vector pRSET PrP23-231. The recombinant vector was acellularly expressed, and then the purified PrP was purified. The purified protein was used to immunize New Zealand white rabbits to prepare its polyclonal antibody and identified by Western-blot. The results were identified by Western-blot, the target band of about 30kD, the successful expression of the protein, and its polyclonal antibody preparation and Western-blot identification, a band of about 30kD, proves the successful preparation of PrP Clone antibody. Conclusion PrP23-231 was successfully expressed and its polyclonal antibody was successfully prepared, which laid the foundation for further study on the function of PrP23.