胸膜肺炎放线杆菌血清7型菌株体内外培养转录差异的研究  

Study on difference of transcription of Actinobacillus pleuropneumoniae serovar 7 strain in vitro and in vivo

作　　者：李萍[1] 樊毅[1] 殷玥[1] 赵军[1] 黄剑波[1] 徐志文[1,2] 朱玲[1,2] 

机构地区：[1]四川农业大学动物医学院动物生物技术中心,四川成都611130 [2]动物疫病与人类健康四川省重点实验室,四川成都611130

出　　处：《中国兽医科学》2017年第3期352-359,共8页Chinese Veterinary Science

基　　金：四川省科技支撑计划项目(2014NZ0043);教育部"长江学者与创新团队发展计划"项目(I RT13083);四川省十三五育种攻关项目(2016NYZ0052);国家"十二五"科技支撑计划(2015BAD12B04-2.3)

摘　　要：为揭示胸膜肺炎放线杆菌血清7型菌株在体内外培养转录的差异,将该菌培养至对数期采用滴鼻的方式攻毒SPF级昆明小鼠,取小鼠肺组织及菌体纯培养物采用Illumina Hi Seq2500高通量测序平台对该菌在体内外培养的转录差异进行转录组测序(RNA-Seq)分析。结果显示,小鼠肺组织中获取该菌的Clean reads与指定参考基因组的比对效率达15.68%,体外培养菌体的比对效率为97.99%;以后者转录组为对照,共筛选出差异表达基因(DEG)333个,其中上调基因113个,下调基因220个,挖掘新基因6个。对DEG进行GO、COG及KEGG等分析可见,DEG主要富集在能量的产生与转化、碳水化合物转运和代谢、无机离子转运与代谢、氨基酸转运与代谢等方面,主要集中在碳代谢、ATP结合蛋白转运蛋白、嘌呤代谢、氨基酸的生物合成、硫代谢等通路上。本研究对胸膜肺炎放线杆菌血清7型菌株在SPF昆明小鼠体内及体外培养转录差异进行了分析,一定程度上揭示了该菌的致病机制,为后续胸膜肺炎放线杆菌血清7型菌株致病机制的研究提供了依据。To reveal the transcriptional differences of Actinobacillus pleuropneumoniae serotype 7 cultured in vivo and in vitro,the bacteria were cultured to a logarithmic phase and intranasally inoculated to SPF Kunming mice.The lung tissue of the infected mouse and pure cultured bacteria were used for transcriptome sequencing（RNA-Seq）,and the differences of transcription of the bacteria that cultured in vivo and in vitro by Illumina were analyzed.The results showed that,the alignment efficiency of the bacterial clean reads from the lung tissue of mouse was 15.68%,compared with the reference genome.As the transcriptome of the latter was the control,a total of 333 differentially expressed genes（DEG） were filtered out, of which 113 were up-regulated and 220 were down regulated,and 6 were new genes.GO,COG and KEGG analysis of DEG showed that these DEG were mainly enriched in＂energy production and conversion＂,＂inorganic ion transport and metabolism＂,＂amino acid transport and metabolism＂,et al.These DEG were also mainly enriched in pathways like carbon metabolism,ABC transporters,purine metabolism,biosynthesis of amino acids,sulfur metabolism,et al.The current study on the transcriptional difference of A. pleuropneumoniae serotype 7 cultured in SPF Kunming mouse revealed the pathogenic mechanism of the bacteria to a certain extent,and provided some bases for future studies on the pathogenic mechanisms.

关 键 词：胸膜肺炎放线杆菌 转录组学 差异基因 

分 类 号：S852.61[农业科学—基础兽医学]