Sean S J Heng, Oliver Y W Chan, Bryan M H Keng, Maurice H T Ling
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引用次数: 29
摘要
基因表达研究中使用的内参基因的表达在大多数情况下是稳定的。然而,许多研究表明,这些基因在实验条件下是不同的。此外,在一个器官中稳定表达的基因可能在其他器官或其他生物体中不稳定表达,这表明需要为每个器官和生物体确定内参基因。本研究旨在鉴定大肠杆菌中稳定表达的基因。分析大肠杆菌MG1655亚株的微阵列数据集和W3110的1个数据集。计算变异系数(COV),并利用NormFinder对3个MG1655组合中4631个共有基因中最低COV的10%进行分析。参与细胞壁合成的葡聚糖生物合成蛋白G (Glucan biosynthesis protein G, mdoG)在MG1655数据集中的加权COV和加权NormFinder稳定性指数最低,同时在菌株W3110数据集中也表现出最稳定的特征,表明mdoG是大肠杆菌K-12的合适内参基因。对39个基因的基因本体超表征分析表明,这些基因在细胞分裂、碳水化合物代谢和蛋白质合成等方面存在过表征,支持了大肠杆菌较短的世代时间。
Glucan Biosynthesis Protein G Is a Suitable Reference Gene in Escherichia coli K-12.
The expressions of reference genes used in gene expression studies are assumed to be stable under most circumstances. However, a number of studies had demonstrated that such genes were found to vary under experimental conditions. In addition, genes that are stably expressed in an organ may not be stably expressed in other organs or other organisms, suggesting the need to identify reference genes for each organ and organism. This study aims at identifying stably expressed genes in Escherichia coli. Microarray datasets from E. coli substrain MG1655 and 1 dataset from W3110 were analysed. Coefficient of variance (COV) of was calculated and 10% of the lowest COV from 4631 genes common in the 3 MG1655 sets were analysed using NormFinder. Glucan biosynthesis protein G (mdoG), which is involved in cell wall synthesis, displayed the lowest weighted COV and weighted NormFinder Stability Index for the MG1655 datasets, while also showing to be the most stable in the dataset for substrain W3110, suggesting that mdoG is a suitable reference gene for E. coli K-12. Gene ontology over-representation analysis on the 39 genes suggested an over-representation of cell division, carbohydrate metabolism, and protein synthesis which supports the short generation time of E. coli.