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


CONCLUSIONS
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.
Mao
Chunhong
C
Virginia Bioinformatics Institute, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA. chmao@vt.edu
Evans
Clive
C
Jensen
Roderick V
RV
Sobral
Bruno Ws
BW
D002874:Chromosome Mapping; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D020869:Gene Expression Profiling; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D016962:Sinorhizobium meliloti / Q000235:genetics*
D004269:DNA, Bacterial