重组利什曼原虫表面糖蛋白GP63的表达生产及其免疫学鉴定

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本文报道应用从国外引进的由硕大利什曼原虫 DNA克隆的原虫细胞表面糖蛋白 GP6 3基因与 p Bluescript M13质粒构建的重组质粒 p AS2 6转化大肠菌 XL1- Blue进行生产性表达及免疫学鉴定的结果。每 2 5 0 m L L B培养物可获取融合蛋白包涵体 5 3mg。表达产物在降解条件下与β半乳糖苷酶分离后的分子量为 5 5 k D。重组 GP6 3及其免疫家兔血清与细菌来源的β半乳糖苷酶有低滴度交叉。与婴儿利什曼原虫 90 1株前鞭毛体及其免疫血清和黑热病人血清 EL ISA反应滴度很高。表明重组 GP6 3抗原可用于黑热病血清学诊断及免疫预防的研究 In this paper, the recombinant plasmid p AS2 6 from protozoal cell surface glycoprotein GP6 3 and p Bluescript M13 cloned from foreign Leishmania DNA was transformed into Escherichia coli XL1-Blue for production and immunological identification the result of. Fusion protein inclusion bodies 5 3 mg were obtained per 250 mL culture. The molecular weight of the expression product after separation from β-galactosidase under the conditions of degradation was 55 kD. Recombinant GP6 3 and its immune rabbit serum and bacterial-derived β-galactosidase low titers cross. Serum ELISA reaction titer was high with 901 strains of promastigotes of Leishmania infantum and its immune and kala-azar. The results showed that the recombinant GP6 3 antigen could be used for the serological diagnosis and immunoprophylaxis of kala-azar
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