基因芯片技术探究肺炎克雷伯菌RcsB的转录谱

Identification of RcsB Transcriptional Profile in Klebsiella pneumoniae by Microarray

肺炎克雷伯菌（Klebsiella pneumoniae,KP）是临床上重要的条件致病菌,主要引起肺炎、泌尿系统疾病、肝脓肿以及菌血症等临床疾病。Rcs B是磷酸信号转导系统中的核心调控子,有证据证明Rcs B可调控部分肠杆菌毒力的表达。本研究利用基因芯片技术筛选出Rcs B调控的靶基因并进行生物信息学分析,同时实时荧光定量PCR技术验证基因芯片结果。通过基因芯片技术共筛选出223个差异基因,其中表达下调基因有132个,表达上调基因有91个。q RT-PCR实验显示,挑选的8个基因的q RT-PCR结果均与芯片结果相符。223个差异基因的COG分析表明Rcs B调控的靶基因主要影响能量的产生和转换,碳、氨基酸的转运和代谢以及细胞壁、外膜的生成等细胞途径。GO富集分析表明Rcs B调控的靶基因主要参与碳水化合物衍生物代谢,酶的催化活性,膜合成等生理生化过程。本研究通过全基因组芯片技术对肺炎克雷伯菌Rcs B的转录谱进行了探究,明确了Rcs B调控的靶基因的相关功能及其调控机制,为通过探究Rcs B的靶基因来进一步深入了解Rcs磷酸信号转导系统对肺炎克雷伯菌毒力的影响奠定了基础。

Klebsiella pneumoniae（K.pneumoniae） is an important opportunistic pathogen in clinical.Rcs B is an essential regulator in Rcs phosphorelay system which could influence the virulence expression of some Enterobacter.The differential expression genes regulated by Rcs B were identified by using microarray technology.Meanwhile,q RT-PCR was used to verify the microarray results.The target genes function annotation was carried out by using COG and GO analysis.A total of 223 differential expression genes were screened out,of which 132 genes were down-regulated and 91 genes were up-regulated.The q RT-PCR results of the 8 genes and the microarray data are consistent.The results of COG analysis indicated the genes regulated by Rcs B mainly belong to energy production and conversion,carbohydrate transport and metabolism,amino acid transport and metabolism,cell wall/membrane/envelope biogenesis.The results of GO analysis showed that the target genes mainly involved in catalytic activity,carbohydrate derivative metabolic process,membrane biogenesis and any other physiological and biochemical processes.In this study,microarray technology was used to explore the genes regulated by Rcs B regulator,and the functions and regulation mechanism of these target genes were indentified,which may provide a basis for investigating the influence of Rcs phosphorelay system on Klebsiella pneumoniae virulence.