National Research Council, Washington, DC 20001, USA.
J Microbiol Methods. 2011 Dec;87(3):320-4. doi: 10.1016/j.mimet.2011.09.008. Epub 2011 Sep 22.
The identification, production, and potential electron conductivity of bacterial extracellular nanofilaments is an area of great study, specifically in Shewanella oneidensis MR-1. While some studies focus on nanofilaments attached to the cellular body, many studies require the removal of these nanofilaments for downstream applications. The removal of nanofilaments from S. oneidensis MR-1 for further study requires not only that the nanofilaments be detached, but also for the cell bodies to remain intact. This is a study to both qualitatively (AFM) and quantitatively (LC/MS-MS) assess several nanofilament shearing methods and determine the optimal procedure. The best method for nanofilament removal, as judged by maximizing extracellular filamentous proteins and minimizing membrane and intracellular proteins, is vortexing a washed cell culture for 10 min.
细菌细胞外纳米丝的鉴定、生产和潜在电子传导性是一个研究热点，特别是在希瓦氏菌属（Shewanella oneidensis）MR-1 中。虽然一些研究集中在附着在细胞体上的纳米丝，但许多研究需要去除这些纳米丝以进行下游应用。为了进一步研究而从希瓦氏菌属（Shewanella oneidensis）MR-1 中去除纳米丝不仅需要将纳米丝分离，还需要保持细胞体完整。本研究旨在定性（AFM）和定量（LC/MS-MS）评估几种纳米丝剪切方法，并确定最佳程序。通过最大限度地增加细胞外丝状蛋白并最小化膜和细胞内蛋白，判断出的最佳纳米丝去除方法是将洗涤后的细胞培养物涡旋 10 分钟。
