LABIOTEC - Biotechnology Laboratory, Department of Biochemistry, Institute of Biology CP6109, University of Campinas - UNICAMP 13083-970, Campinas, São Paulo, Brazil.
J Nanobiotechnology. 2014 Apr 16;12:14. doi: 10.1186/1477-3155-12-14.
H. influenzae is a natural competent bacterium that can uptake DNA from the environment and recombine into bacterial genome. The outbreaks of Brazilian purpuric fever, heavily polluted areas of a different H. influenzae biogroup - aegyptius - as well as gene transference between Neisseria meningitis make the transformation process an important evolutionary factor. This work studied the horizontal transference of the ompP2 gene from a multiresistant strain of H. influenzae 07 (NTHi), under the influence of graphene oxide nanoparticles in order to mimic an atmosphere rich in suspended particles and this way verify if the CFU transformants number was increased.
In this article the gene ompP2 was transformed into different strains of H. influenzae mediated or not by graphene oxide nanoparticles in suspension, followed by the adhesion tests in Hec-1B (human endometrium adenocarcinoma) and A549 (pulmonary epithelial carcinoma) cells lines. The transformation frequency and the adhesion capacity were determined in all the mutants to which the ompP2 gene was transferred and compared to their wild type strains.
The nanoparticles increased the transformation ratio of one particular strain isolated from a pneumonia case. The adhesion patterns to A549 and Hec1b cell lines of these mutated bacteria has their capacity increased when compared to the wild type.
Graphene oxide nanoparticles aid the transformation process, helping to increase the number of CFUs, and the mutants generated with the ompP2 gene from a H. influenzae resistant strain not only present a chloramphenicol resistance but also have an increased adherence patterns in A549 and Hec1B cell lines.
流感嗜血杆菌是一种天然感受态细菌，能够从环境中摄取 DNA 并将其重组到细菌基因组中。巴西紫癜热的爆发、不同生物群组的流感嗜血杆菌（埃及生物群组）在污染严重的地区的爆发，以及脑膜炎奈瑟菌之间的基因转移，使得转化过程成为一个重要的进化因素。本研究旨在模拟富含悬浮颗粒的大气环境，研究氧化石墨烯纳米粒子对 H. influenzae 07（NTHi）多耐药株 ompP2 基因的水平转移，以验证 CFU 转化体数量是否增加。
在本文中，我们将 ompP2 基因转化为不同的流感嗜血杆菌菌株，或通过悬浮的氧化石墨烯纳米粒子介导，然后在 Hec-1B（人子宫内膜腺癌）和 A549（肺上皮癌细胞）细胞系中进行黏附试验。在所有转导 ompP2 基因的突变体中，确定了转化频率和黏附能力，并将其与野生型菌株进行比较。
纳米粒子增加了从肺炎病例中分离的一株特定菌株的转化比例。与野生型相比，这些突变菌对 A549 和 Hec1b 细胞系的黏附模式的黏附能力增强。
氧化石墨烯纳米粒子有助于转化过程，有助于增加 CFU 的数量，并且从具有耐氯霉素的流感嗜血杆菌抗性株产生的带有 ompP2 基因的突变体不仅具有氯霉素抗性，而且在 A549 和 Hec1B 细胞系中的黏附模式增加。
