氟苯尼考胁迫下弗氏柠檬酸杆菌转录组测序研究

doi:10.16431/j.cnki.1671-7236.2019.11.005

摘要/Abstract

摘要： 为了探究弗氏柠檬酸杆菌的耐药机制，试验使用氟苯尼考为诱导药物，提取正常培养与400 μg/mL氟苯尼考胁迫下的菌株RNA，利用转录组测序技术对菌株间基因表达水平与差异表达基因进行了分析。结果显示，每个样品平均产出1.31 Gb数据，与参考基因组的平均比对率为67.01%。对样品间差异表达基因进行检测，共筛选出2 019个显著差异表达基因，其中上调963个，下调1 056个，并发现emrA和emrB 2个基因。KEGG Pathway分析中，富集到代谢途径的显著差异表达基因最多，占全部的72%，碳水化合物代谢、能量代谢、氨基酸代谢、膜转运等通路明显富集。GO功能分析中，生物过程分布的显著差异基因最多，占全部的46%。显著差异表达基因主要涉及到代谢进程、催化活性、结合。本试验结果表明，弗氏柠檬酸杆菌耐药性的产生可能与细胞外排泵介导的主动外排作用、靶位改变、酶的水解作用、细胞膜通透性改变及生物膜的形成有关。

关键词: 弗氏柠檬酸杆菌; 氟苯尼考; 转录组测序; 耐药性

Abstract: In order to explore the resistance mechanism of Citrobacter freundii,florfenicol was used as an inducer,RNA was extracted from normal cultured strains and strains under flufenicol stress at 400 μg/mL.Gene expression levels and differentially expressed genes between strains were analyzed by RNA sequencing (RNA-Seq).The results showed that the average output of each sample was 1.31 Gb.The mean alignment rate with reference genome was 67.01%.Through the detection of differentially expressed genes among samples,a total of 2 019 significantly differentially expressed genes were screened,among which 963 genes were up-regulated and 1 056 genes were down-regulated,and 2 genes of emrA and emrB were found.KEGG Pathway analysis showed that the most differentially expressed genes were enriched in metabolism,accounting for 72% of the total.Carbohydrate metabolism,energy metabolism,amino acid metabolism,membrane transport and other pathways were significantly enriched.GO functional analysis showed that the distribution of significantly different genes in biological processes was the largest,accounting for 46% of all genes.Significant differentially expressed genes mainly involved metabolic process,catalytic activity and binding.It was suggested that the resistance of Citrobacter freundii might be related to active efflux mediated by extracellular pump,target change,enzyme hydrolysis,membrane permeability change and biofilm formation.

Key words: Citrobacter fredii; fluphenicol; RNA-Seq; drug resistance

中图分类号:

S852.61

韦盘秋, 黄杰, 韩书煜, 罗福广, 黎娅, 易弋. 氟苯尼考胁迫下弗氏柠檬酸杆菌转录组测序研究[J]. 中国畜牧兽医, 2019, 46(11): 3172-3180.

WEI Panqiu, HUANG Jie, HAN Shuyu, LUO Fuguang, LI Ya, YI Yi. RNA Sequencing of Citrobacter fredii Under Fluphenicol Stress[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(11): 3172-3180.

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