摘要
为探究内化素G在单增李斯特菌(LM)致病性及生物学特性中的作用,以单增李斯特菌ATCC19111标准株为模板,扩增出用于缺失内化素G(Inl G)的上、下游同源臂。应用同源重组将上、下游同源臂融合扩增得到LM-ΔInl G缺失株,然后比较分析标准株与缺失株的生长特性、对细胞的侵袭能力及转录组测序结果。结果显示,LM-ΔInl G的生长速度与LM相比无显著变化,Inl G不参与温度调节,不影响细菌对酸碱的耐受性;细胞黏附侵袭试验表明,LM-ΔInl G增强了LM对细胞的内化作用(P<0.01);GO富集分析显示,显著差异表达基因主要富集在裂解酶活性、小分子结合和氮化合物代谢过程;KEGG pathway分析,显著差异表达基因主要参与群体感应、丙酸代谢,缬氨酸、亮氨酸和异亮氨酸降解、甘油磷脂代谢和丁酸代谢等22个代谢通路。表明Inl G与LM的毒力有关,缺失Inl G可以加强对细胞的内化作用,为阐明LM的致病机理提供了科学依据。
To explore the role of internalin G in the pathogenicity and biological characteristics of Listeria monocytogenes(LM),the standard strain of LM ATCC19111 was used as a template to amplify the upstream and downstream homologous arms for the deletion of internalin G(Inl G).The two homology arms were fused and amplified by homologous recombination to obtain LM-ΔInl G deletion strain,and then compared and analyzed the growth characteristics,cell invasion ability and transcriptome sequencing of the standard strain and the mutant strain.The results showed that the growth rate of LM-ΔInl G had no significant change compared with LM.Inl G gene did not participate in temperature regulation and did not affect the tolerance of bacteria to acid and alkali.Cell adhesion and invasion test showed that LM-ΔInl G enhanced the internalization of LM on cells(P<0.01).GO enrichment analysis showed that the differentially expressed genes were mainly enriched in lyase activity,small molecule binding and nitrogen compound metabolic process.KEGG pathway analysis showed that the differentially expressed genes were mainly involved in 22 pathways including quorum sensing,propanoate metabolism,valine,leucine and isoleucine degradation,glycerophospholipid metabolism and butanoate metabolism.These data indicated that Inl G is related to the virulence of LM,and the deletion of Inl G can strengthen the internalization of cells,which provides a scientific basis for elucidating the pathogenesis of LM.
作者
史文静
刘圆园
王子坚
王学俭
齐玉梅
田常青
苟惠天
薛慧文
SHI Wen-jing;LIU Yuan-yuan;WANG Zi-jian;WANG Xue-jian;QI Yu-mei;TIAN Chang-qing;GOU Hui-tian;XUE Hui-wen(Gansu Agricultural University,Lanzhou,Gansu 730070,China;Gansu Animal Disease Prevention and Control Center,Lanzhou,Gansu 730199,China;Xigu Center for Animal Disease Control and Prevention,Lanzhou,Gansu 730060,China)
出处
《动物医学进展》
北大核心
2023年第10期56-62,共7页
Progress In Veterinary Medicine
基金
国家自然科学基金项目(31960726,32060822)
国家重点研发计划项目(2019YFC1605705)
甘肃省重点研发计划项目(20YF8FA136)
甘肃农业大学青年导师基金(GAU-QDFC-2020-010)。
关键词
单增李斯特菌
内化素G
同源重组
缺失株
转录组
Listeria monocytogenes
internalin G
homologous recombination
mutant strain
transcriptome
