Identification of new genes in Sinorhizobium meliloti using the Genome Sequencer FLX system

BMC Microbiol. 2008 May 2:8:72. doi: 10.1186/1471-2180-8-72.

Abstract

Background: Sinorhizobium 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.

Results: Our 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.

Conclusion: Our 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromosome Mapping
  • DNA, Bacterial / analysis
  • DNA, Bacterial / genetics
  • Gene Expression Profiling*
  • Genes, Bacterial
  • Genome, Bacterial*
  • Molecular Sequence Data
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Sinorhizobium meliloti / genetics*

Substances

  • DNA, Bacterial