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在肉汤和琼脂中生长的副猪嗜血杆菌的转录组差异。

Transcriptomic differences noted in Glaesserella parasuis between growth in broth and on agar.

机构信息

Virus and Prion Research Unit, National Animal Disease Center, ARS, USDA, Ames, Iowa, United States of America.

Oak Ridge Institute for Science and Education, Oak Ridge, Tennessee, United States of America.

出版信息

PLoS One. 2019 Aug 6;14(8):e0220365. doi: 10.1371/journal.pone.0220365. eCollection 2019.

Abstract

Glaesserella parasuis is the cause of Glӓsser's disease in pigs and is a significant contributor to post-weaning mortality in the swine industry. Prevention of G. parasuis disease relies primarily on bacterin vaccines, which have shown good homologous protection and variable heterologous protection. Bacterin production involves large scale growth of the bacteria and proteins produced during the proliferation phase of production become important antigens that stimulate the immune response. In order to evaluate genes activated during G. parasuis growth on different media substrates, the transcriptome of broth and agar grown G. parasuis strain 29755 were sequenced and compared. The transcription of most purported virulence genes were comparable between broth and agar grown G. parasuis; however, four virulence-associated genes, including ompA and vapD, had elevated expression under agar growth, while six virulence-associate genes had elevated expression during broth growth, including several protease genes. Additionally, there were metabolic shifts toward increased protein and lipid production and increased cellular division in broth grown G. parasuis. The results contribute to the understanding of how growth substrate alters gene transcription and protein expression, which may impact vaccine efficacy if immunogens important to the protective immune response are not produced under specific in vitro conditions. While the results of this work are unable to fully elucidate which growth medium presents a transcriptome more representative of in vivo samples or best suited for bacterin production, it forms a foundation that can be used for future comparisons and provides a better understanding of the metabolic differences in broth and agar grown bacteria.

摘要

副猪嗜血杆菌是导致猪格拉瑟氏病的病原体,也是猪养殖业中小猪断奶后死亡的主要原因之一。副猪嗜血杆菌病的预防主要依赖于菌苗,菌苗对同源菌株具有良好的保护效果,对异源菌株具有不同程度的保护效果。菌苗的生产涉及到细菌的大规模生长,在生产的增殖阶段产生的蛋白质成为重要的抗原,刺激免疫反应。为了评估不同培养基基质上副猪嗜血杆菌生长过程中激活的基因,对肉汤和琼脂培养的副猪嗜血杆菌 29755 菌株的转录组进行了测序和比较。肉汤和琼脂培养的副猪嗜血杆菌中大多数假定的毒力基因的转录水平相当;然而,有 4 个与毒力相关的基因,包括 ompA 和 vapD,在琼脂生长时表达上调,而 6 个与毒力相关的基因在肉汤生长时表达上调,包括几个蛋白酶基因。此外,肉汤培养的副猪嗜血杆菌中还存在向蛋白质和脂质生产增加以及细胞分裂增加的代谢转变。这些结果有助于了解生长基质如何改变基因转录和蛋白质表达,如果对保护性免疫反应重要的免疫原在特定的体外条件下不能产生,可能会影响疫苗的效果。虽然这项工作的结果还不能完全阐明哪种生长培养基更能代表体内样本的转录组,或者最适合菌苗生产,但它为未来的比较提供了基础,并更好地理解了肉汤和琼脂培养细菌之间的代谢差异。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de0b/6684057/b037be96b1d0/pone.0220365.g001.jpg

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