らい菌のα抗原類似蛋白質(α2抗原)遺伝子のクロ-ニングと大腸菌中での大量発現

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I have constructed the genomic library of M. leprae Thai 53 strain, and cloned the α antigen like protein gene by plaque hybridization method by using M. leprae α antigenDNA fragment as probe which was characterized in the previous study, I have termed it as α2 antigen gene. The α2 antigen gene has been characterized by sequencing. By comparing the deduced amino acid sequence of α and α2 antigen with 85 complex antigen of other mycobacteria. Ihave found the higher homology between α2 antigen and 85A antigen and between a antigen and 85B antigen. We have constructed the over expression system of M. leprae α and α2 antigengene in E. coli using vector pMALc-RI. Recombinant α and α2 antigen has been purified byamylose column chromatography at the purity of more than 95%. More than 6 mg and more than 10 mg of recombinant α and α2 antigen has been obtained from 200 ml of liquid culture, respectively. ELISA tests have been performed with the sera of leprosy patient and healthy control against the recombinant α and α2 antigens. The antibody titers in sera of leprosy patient against the two kinds of antigens were all much higher than healthy controls. The antibody titer against the α2 antigen was higher than that against α antigen. Recombinant α and α2 antigens in this study could be used as a new specific antigen for serodiagnosis of leprosy.
日本感染症学会の論文