Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA.
Genome Res. 2011 Feb;21(2):315-24. doi: 10.1101/gr.107854.110. Epub 2010 Dec 22.
RNA-seq was used to generate an extensive map of the Drosophila melanogaster transcriptome by broad sampling of 10 developmental stages. In total, 142.2 million uniquely mapped 64-100-bp paired-end reads were generated on the Illumina GA II yielding 356× sequencing coverage. More than 95% of FlyBase genes and 90% of splicing junctions were observed. Modifications to 30% of FlyBase gene models were made by extension of untranslated regions, inclusion of novel exons, and identification of novel splicing events. A total of 319 novel transcripts were identified, representing a 2% increase over the current annotation. Alternate splicing was observed in 31% of D. melanogaster genes, a 38% increase over previous estimations, but significantly less than that observed in higher organisms. Much of this splicing is subtle such as tandem alternate splice sites.
RNA-seq 技术通过广泛采样 10 个发育阶段,生成了一个详尽的黑腹果蝇转录组图谱。在 Illumina GA II 上共生成了 1.422 亿个独特映射的 64-100-bp 配对末端读取,测序覆盖率达到 356×。超过 95%的 FlyBase 基因和 90%的剪接接头都被观察到。通过延长非翻译区、包含新外显子和鉴定新的剪接事件,对 30%的 FlyBase 基因模型进行了修改。总共鉴定出 319 个新的转录本,比当前注释增加了 2%。在 31%的黑腹果蝇基因中观察到可变剪接,比之前的估计增加了 38%,但明显低于在高等生物中观察到的水平。这种剪接中有很多是细微的,例如串联的可变剪接位点。