Comerci D J, Pollevick G D, Vigliocco A M, Frasch A C, Ugalde R A
Instituto de Investigaciones Biotecnologicas, Universidad Nacional de General San Martin, INTI, San Martin, Provincia de Buenos Aires, Argentina.
Infect Immun. 1998 Aug;66(8):3862-6. doi: 10.1128/IAI.66.8.3862-3866.1998.
A vector for the expression of foreign antigens in the vaccine strain Brucella abortus S19 was developed by using a DNA fragment containing the regulatory sequences and the signal peptide of the Brucella bcsp31 gene. This fragment was cloned in broad-host-range plasmid pBBR4MCS, resulting in plasmid pBEV. As a reporter protein, a repetitive antigen of Trypanosoma cruzi was used. The recombinant fusion protein is stably expressed and secreted into the Brucella periplasmic space, inducing a good antibody response against the T. cruzi antigen. The expression of the repetitive antigen in Brucella neither altered its growth pattern nor generated a toxic or lethal effect during experimental infection. The application of this strategy for the generation of live recombinant vaccines and the tagging of B. abortus S19 vaccine is discussed. This is the first time that a recombinant protein has been expressed in the periplasm of brucellae.
通过使用包含布鲁氏菌bcsp31基因调控序列和信号肽的DNA片段，构建了一种用于在疫苗株布鲁氏菌流产菌S19中表达外源抗原的载体。该片段克隆于广宿主范围质粒pBBR4MCS中，得到质粒pBEV。使用克氏锥虫的一种重复抗原作为报告蛋白。重组融合蛋白在布鲁氏菌周质空间中稳定表达并分泌，诱导产生针对克氏锥虫抗原的良好抗体反应。克氏锥虫重复抗原在布鲁氏菌中的表达既未改变其生长模式，在实验感染期间也未产生毒性或致死效应。讨论了该策略在生产活重组疫苗和布鲁氏菌流产菌S19疫苗标记方面的应用。这是首次在布鲁氏菌周质中表达重组蛋白。
