Kobatake E, Iwai T, Ikariyama Y, Aizawa M
Department of Bioengineering, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Anal Biochem. 1993 Feb 1;208(2):300-5. doi: 10.1006/abio.1993.1050.
Protein A and firefly luciferase were genetically fused and the resulting fusion protein was applied to a bioluminescent immunoassay. The gene fusion plasmid, pMALU2, was constructed by inserting the structural gene of luciferase into a protein A expression vector, and was expressed in Escherichia coli. The resulting fusion protein of molecular weight 91 kDa retained not only the enzymatic activity of luciferase but also the binding capability of protein A to the Fc region of immunoglobulin G (IgG). The bioluminescent immunoassay was performed with the fusion protein and human IgG was determined in the concentration range from 10(-3) to 10(-7) g/ml.
将蛋白A与萤火虫荧光素酶进行基因融合，并将所得融合蛋白应用于生物发光免疫测定。通过将荧光素酶的结构基因插入蛋白A表达载体构建基因融合质粒pMALU2，并在大肠杆菌中表达。所得分子量为91 kDa的融合蛋白不仅保留了荧光素酶的酶活性，还保留了蛋白A与免疫球蛋白G（IgG）Fc区的结合能力。用该融合蛋白进行生物发光免疫测定，测定人IgG的浓度范围为10(-3)至10(-7) g/ml。
