基于RNA-Seq的羊种布鲁氏菌新转录本与非编码RNA鉴定  

Identification of novel transcripts and sRNA of Brucella melitensis by RNA-Seq

作　　者：郭英飞[1,2] 王玉飞[3] 龚春丽[1] 杨明娟[1] 袁久云[1] 庄妤冰 柯跃华[1] 杜昕颖[1] 汪舟佳[1] 陈泽良[1] 

机构地区：[1]军事医学科学院疾病预防控制所,北京100071 [2]空军司令部门诊部,北京100843 [3]武警总医院检验科,北京100069

出　　处：《中国人兽共患病学报》2015年第3期216-221,共6页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(81171530);北京市科技新星资助项目(Z131102000413062);北京市自然基金项目(6122030)~~

摘　　要：目的对羊种布鲁氏菌的转录本进行测序,鉴定基因组中新的转录本和非编码RNA。方法提取羊布鲁氏菌的总RNA,去除rRNA后连接接头逆转录成cDNA,PCR扩增后进行测序,以羊布鲁氏菌16M的基因组序列为参照对测序得到的reads进行比对作图,通过生物信息学方法进行新转录本和非编码RNA的鉴定。结果测序数据分析显示,reads在基因组上覆盖度较好。与现有注释基因相比,有773个基因的5′或3′端在基因组的原有位置基础上发生了延伸,并发现了16个新的转录本。根据测序结果,共鉴定出241条候选的非编码RNA(sRNA),进一步的RT-PCR验证结果显示,预测的sRNA在体外条件下有表达。结论布鲁氏菌基因组中存在除了预测以外的新的转录本,布鲁氏菌中存在非编码RNA且在不同条件下有差异表达。To identify novel transcripts and sRNA in genome of B .melitensis by transcriptome sequencing ,total RNA were extracted from B .melitensis culture and rRNA were removed .After the addition of adaptor ,RNA was reversely transcribed into cDNA ,which were then subjected to PCR amplification and sequencing .The generated reads were mapped to genome se‐quence of B .melitensis strain 16M .With the mapping results ,novel transcripts and sRNA were identified by bioinformatics methods .Sequencing results analysis showed that genome sequence was covered with the reads with good quality .A total of 773 genes were extended in their 5′and/or 3′ends of their original locations .Sixteen novel transcripts and 241 sRNAs candi‐dates were identified .RT‐PCR showed that some of the sRNAs were differentially expressed under stress conditions .In B . melitensis genome ,there is novel transcript which is not predicted .The sRNA does exist in B .melitensis and were expressed under different conditions .

关 键 词：布鲁氏菌 RNA-SEQ 转录组 SRNA 

分 类 号：R378.5[医药卫生—病原生物学]