Crosby Francy L, Brayton Kelly A, Magunda Forgivemore, Munderloh Ulrike G, Kelley Karen L, Barbet Anthony F
College of Veterinary Medicine, University of Florida, Department of Infectious Diseases and Pathology, Gainesville, Florida, USA
College of Veterinary Medicine, Washington State University, Department of Veterinary Microbiology and Pathology, Pullman, Washington, USA.
Appl Environ Microbiol. 2015 Mar;81(6):2206-14. doi: 10.1128/AEM.03241-14. Epub 2015 Jan 16.
Anaplasma marginale is the causative agent of anaplasmosis in cattle. Transposon mutagenesis of this pathogen using the Himar1 system resulted in the isolation of an omp10 operon insertional mutant referred to as the omp10::himar1 mutant. The work presented here evaluated if this mutant had morphological and/or growth rate defects compared to wild-type A. marginale. Results showed that the morphology, developmental cycle, and growth in tick and mammalian cell cultures are similar for the mutant and the wild type. Tick transmission experiments established that tick infection levels with the mutant were similar to those with wild-type A. marginale and that infected ticks successfully infected cattle. However, this mutant exhibited reduced infectivity and growth in cattle. The possibility of transforming A. marginale by transposon mutagenesis coupled with in vitro and in vivo assessment of altered phenotypes can aid in the identification of genes associated with virulence. The isolation of deliberately attenuated organisms that can be evaluated in their natural biological system is an important advance for the rational design of vaccines against this species.
边缘无形体是牛无形体病的病原体。使用Himar1系统对该病原体进行转座子诱变,导致分离出一个被称为omp10::himar1突变体的omp10操纵子插入突变体。本文的研究评估了该突变体与野生型边缘无形体相比是否存在形态和/或生长速率缺陷。结果表明,该突变体与野生型在形态、发育周期以及在蜱和哺乳动物细胞培养物中的生长情况相似。蜱传播实验证实,突变体在蜱中的感染水平与野生型边缘无形体相似,且感染的蜱能够成功感染牛。然而,该突变体在牛体内的感染性和生长能力有所降低。通过转座子诱变转化边缘无形体,并结合对改变表型的体外和体内评估,有助于鉴定与毒力相关的基因。分离出可在其自然生物系统中进行评估的经过特意减毒的生物体,是针对该物种合理设计疫苗的一项重要进展。