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为研究迟缓爱德华菌菌毛蛋白PILI的功能和免疫原性,本研究利用PCR方法从迟缓爱德华菌基因组中克隆出pili基因,并将其构建到pET-32a原核表达载体上,重组载体转化BL21大肠杆菌诱导重组蛋白表达,通过SDSPAGE和Western blot方法验证重组蛋白表达。用亲和层析方法纯化蛋白,纯化的蛋白免疫小鼠,所得的多抗血清能够特异性识别迟缓爱德华菌PILI蛋白,为研究PILI蛋白奠定基础。
In order to study the function and immunogenicity of delayed PIH PILI, the pili gene was cloned from the genome of L. edwardsiana by PCR and cloned into pET-32a prokaryotic expression vector. The recombinant vector was transformed into BL21 large intestine Bacillus coli induced the expression of recombinant protein, and the recombinant protein was verified by SDSPAGE and Western blot. The purified protein was purified by affinity chromatography and the purified protein was used to immunize mice. The obtained polyclonal antiserum specifically recognized PILI protein, which laid the foundation for the study of PILI protein.