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链霉菌作为重组生产结核分枝杆菌蛋白的宿主。

Streptomyces as host for recombinant production of Mycobacterium tuberculosis proteins.

作者信息

Vallin Carlos, Ramos Astrid, Pimienta Elsa, Rodríguez Caridad, Hernández Tairí, Hernández Ivones, Del Sol Ricardo, Rosabal Grisel, Van Mellaert Lieve, Anné Jozef

机构信息

Department of Biomedical Research, Center of Pharmaceutical Chemistry, Atabey, Playa, Havana, Cuba.

出版信息

Tuberculosis (Edinb). 2006 May-Jul;86(3-4):198-202. doi: 10.1016/j.tube.2006.02.003. Epub 2006 Apr 27.

Abstract

The 45/47 kDa APA protein (Rv1860) of Mycobacterium tuberculosis was produced by Streptomyces lividans. The recombinant protein could be recovered from the culture medium of an S. lividans clone containing the apa gene under control of the promoter and signal sequence of the Streptomyces coelicolor agarase gene. The recombinant protein production was further scaled-up using fermentation conditions. The APA protein was subsequently purified from the culture supernatant by means of immunochromatography. About 80 mg of recombinant protein were obtained per liter of culture media. In vivo tests with the APA protein purified from S. lividans TK24/pRGAPA1 revealed that the recombinant protein was antigenic and could induce high titers of specific antibodies in the mouse biological model. Results obtained concerning heterologous production of APA, its immunogenic and antigenic capacity, demonstrated the potential of S. lividans as a valuable host for the production of recombinant proteins from M. tuberculosis.

摘要

结核分枝杆菌的45/47 kDa APA蛋白(Rv1860)由淡紫链霉菌产生。重组蛋白可从含有apa基因的淡紫链霉菌克隆的培养基中回收,该基因受天蓝色链霉菌琼脂酶基因的启动子和信号序列控制。利用发酵条件进一步扩大重组蛋白的生产规模。随后通过免疫色谱法从培养上清液中纯化APA蛋白。每升培养基可获得约80 mg重组蛋白。对从淡紫链霉菌TK24/pRGAPA1中纯化的APA蛋白进行的体内试验表明,该重组蛋白具有抗原性,可在小鼠生物学模型中诱导产生高滴度的特异性抗体。关于APA的异源生产及其免疫原性和抗原性的研究结果,证明了淡紫链霉菌作为生产结核分枝杆菌重组蛋白的有价值宿主的潜力。

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