从表达菌培养液中提取鼠疫菌重组F1抗原

Extracting recombinant F1 antigen of Yersinia pestis from culture medium of expression bacteria

目的从表达鼠疫耶尔森氏菌F1蛋白的诱导培养液中提取纯化重组F1并检测其免疫反应性。方法收集诱导培养后离心去除菌体的培养液,采用硫酸铵沉淀法和PEG浓缩法提取F1蛋白,产物经镍离子金属螯合亲和层析纯化,免疫印迹(Western blot)鉴定其免疫反应性。结果硫酸铵沉淀法和PEG浓缩法均能从培养液中提取到重组F1蛋白,产物经纯化后可与鼠疫疫苗株免疫兔血清和既往鼠疫患者血清发生特异性结合反应。结论该表达菌表达效能较高,在诱导培养液中存在大量具有免疫学活性的重组F1蛋白。

Objective To extract purified recombinant F1 antigen from inducing medium expressing Yersiniapestis F1 pro- tein and detect its immune response. Methods The inducing medium removed bacteria by centrifugation were collected. The F 1 proteins of medium were extracted using the ammonium sulfate precipitation and PEG concentration method. The products were purified by immobilized metal Nickel ion affinity chromatography and their immune responses were identified by Western blot. Results The recombinant F1 protein was extracted using both the ammonium sulfate precipitation and PEG concentration method. A specific binding reaction occurred between the purified products and sera of rabbit immunized with plague vaccine and sera of patients suffered from plague. Conclusions The expression bacteria have high expressing efficiency. A great deal of the recombinant F 1 protein with immune response can be extracted in inducing medium.