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目的 :利用大肠杆菌中表达的人Bit1(hBit1)蛋白 ,制备兔抗人Bit1抗血清并对其进行纯化。方法 :在大肠杆菌中诱导表达人Bit1融合蛋白。以其免疫家兔 ,制备兔抗人Bit1抗血清 ,用非特异性去除法纯化抗体 ,并以Westernblot对所纯化的兔抗人Bit1抗血清进行鉴定。结果 :大肠杆菌中表达的人Bit1蛋白占菌体总蛋白的 4 0 %。用纯化的抗血清可特异地检出大肠杆菌表达的人Bit融合蛋白。结论 :人Bit1融合蛋白在大肠杆菌中得到高效表达 ,并成功地制备并纯化了兔抗人Bit1抗血清
OBJECTIVE: To prepare and purify rabbit anti-human Bit1 antiserum using human Bit1 (hBit1) protein expressed in Escherichia coli. Methods: Human Bit1 fusion protein was induced in Escherichia coli. Rabbit anti-human Bit1 antiserum was prepared by immunizing rabbits. Antibody was purified by nonspecific removal method and the purified rabbit anti-human Bit1 antiserum was identified by Western blot. Results: Bit1 protein expressed in E. coli accounted for 40% of total bacterial proteins. E. coli-expressed human Bit fusion protein can be specifically detected with the purified antiserum. Conclusion: The human Bit1 fusion protein is highly expressed in E. coli and successfully prepared and purified rabbit anti-human Bit1 antiserum