何进 微生物功能基因组学之转录组学.pptxVIP

微生物功能基因组学之转录组学1微生物转录组学的原理与应用何进mailto:hejin@mail.hzau.edu.cnhejin@mail.hzau.edu.c题?转录transcription 、转录本transcript、转录组transcriptome 、转录组学transcriptomicsRNA的种类、研究方法及主要原理；RNA-seq的基本原理及主要流程。RNA-seq在微生物学研究中有哪些应用？什么是链特异性RNA-seq (ssRNA-Seq)？RNA-seq为什么要除去rRNA，如何除去rRNA?什么是5’-非翻译区5’-UTR？什么是核糖开关riboswitch？核糖开关riboswitch是如何调节基因表达的？什么是反义RNA (Antisense RNA，asRNA)？什么是small RNA (sRNA)? 什么是non-coding RNA (ncRNA)?基础知识Transcription是遗传信息从DNA流向RNA的过程。即以双链DNA中的确定的一条链(模板链用于转录，编码链不用于转录)为模板，以ATP、CTP、GTP、UTP四种核苷三磷酸为原料，在RNA聚合酶催化下合成RNA的过程。 启动子序列 转录因子 RNAP核心酶 σ因子RNAP CoreTranscriptionalfactorσ factor(Haugen et al., Nat Rev Microbiol. 2008, 6(7):507-519. Key points:RNA polymerase slides along the DNA, creating an open complex as it moves.The template strand is used to make a complementary copy of RNA as an RNA–DNA hybrid.The RNA is synthesized in a 5′ to 3′ direction using ribonucleoside triphosphates as precursors. Pyrophosphate is released (not shown).The complementarity rule is the same as the AT/GC rule except that U is substituted for T in the RNA.E. coli (fluorescent image) 基础知识细菌中的RNA分子Volume occupied by RNAs in E. coli cytoplasmProtein-coding RNAstotal RNA ?6%rRNA ?5%tRNA ?0.8%mRNA others ?0.2%mRNANon-coding RNAsrRNA (23S, 16S, 5S)tRNA6S RNAtmRNAsmall RNAanti-sense RNAinternal RNA……基础知识5’-UTRCDSCDSmRNAmRNAantisense RNAinternal RNAsmall RNA基础知识mRNAsThese protein-coding RNA species are transcribed from the coding DNA strand and contain an ORF, a 5? UTR and a 3? UTR.rRNAsRibosomal RNAs are non-coding RNAs that form the nucleic acid component of the ribosome; in bacteria, there are three rRNA species: the 16S rRNA, the 23S rRNA and the 5S rRNA.tRNAs（有无代谢需要tRNA运载aa）These non-coding adaptor-like RNA molecules carry amino acids to the ribosome and facilitate protein synthesis by complementary base pairing with the codons in the mRNA template.tmRNAsRNAs Small RNAs (sRNAs) are stable RNAs that are typically less than 250 nucleotides in length and have a defined structure. These RNAs