Abstract
BackgroundSinorhizobium meliloti is an agriculturally important model symbiont. There is an ongoing need to update and improve its genome annotation. In this study, we used a high-throughput pyrosequencing approach to sequence the transcriptome of S. meliloti, and search for new bacterial genes missed in the previous genome annotation. This is the first report of sequencing a bacterial transcriptome using the pyrosequencing technology.ResultsOur pilot sequencing run generated 19,005 reads with an average length of 136 nucleotides per read. From these data, we identified 20 new genes. These new gene transcripts were confirmed by RT-PCR and their possible functions were analyzed.ConclusionOur results indicate that high-throughput sequence analysis of bacterial transcriptomes is feasible and next-generation sequencing technologies will greatly facilitate the discovery of new genes and improve genome annotation.
Highlights
Sinorhizobium meliloti is an agriculturally important model symbiont
Small gene size and lack of BLAST hits may be the reasons that the predicted new gene candidates were missed in the original genome annotation process
Our study demonstrated that there are many genes missed in the initial genome annotation and it is useful to have large-scale transcriptome analysis to reveal these genes
Summary
Sinorhizobium meliloti is an agriculturally important model symbiont. We used a highthroughput pyrosequencing approach to sequence the transcriptome of S. meliloti, and search for new bacterial genes missed in the previous genome annotation. This is the first report of sequencing a bacterial transcriptome using the pyrosequencing technology. Sinorhizobium meliloti is a micro-symbiont associated with legume plants. Because of its agricultural and ecological importance, S. meliloti has been extensively studied as a model symbiont. The S. meliloti 1021 genome sequence and the initial annotation of the genome were completed in 2001 [1,2,3,4].
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