57º Congresso Brasileiro de Genética Resumos do 57º Congresso Brasileiro de Genética • 30 de agosto a 2 de setembro de 2011 Centro de Convenções do Hotel Monte Real Resort • Águas de Lindóia • SP • Brasil www.sbg.org.br - ISBN 978-85-89109-06-2 125 Transcriptome analysis – Comparing primitively social and social bees Marco Antonio, DS1*; Cristino, AS2; Hartfelder, K3 1 Departamento de Genética, Faculdade de Medicina de Ribeirão Preto – Universidade de São Paulo, Ribeirão Preto email: [email protected] 2 The Queensland Brain Institute, The University of Queensland, St Lucia, Brisbane, Australia, 3 Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto – Universidade de São Paulo, Ribeirão Preto Keywords: Transcriptome, Bee, Social bees, Inseticide sensitivity, Gene family Expressed Sequence Tags (ESTs) are fragments of cDNA between 200-800 bp. They are generated from mRNA through reverse transcription and are very usefull for gene identiﬁcation and prediction. Futhermore, through BLAST analysis it’s possible to associate predicted functions to each EST. The clusterization of ESTs based in annotated genes improves prediction, and unclusterized ESTs can represent speciﬁc genes of the species. Adult workers of Melipona quadrifasciata anthidioides and Frieseomelitta varia were collected when returning from ﬂights and insight nest. Adult females of Euglossa cordata and Eulaema nigrita were collected in the ﬁeld when visiting ﬂowers. The bees were immediately frozen in liquid nitrogen. The brains of 15-30 bees were dissected and total RNA extracted in TRIzol. Sequencing was done on a Roche 454 platform. After validation of ESTs quality they were submited to CAP3 analysis generating contigs and singlets. These sequences were used to assemble the database of each specie. BLAST algorithm were used for alignment analysis with cutoff E-value<10-5 and gap/penalities default. Data mining algorithms were especially designed for the transcriptomes using Shell script and Python (v.2.4 and above). The Apis mellifera genome was chosen as anchor due to phylogenetic proximity and a completely sequenced annotated genome. The dbEST of NCBI encompasses 79,179 Apis mellifera ESTs, of a normalized brain library.For each transcriptome a “best hit” list was created using blastn analysis against Apis mellifera (Ofﬁcial Gene Set 2). A second list was created through blastx searches using the ﬁrst list against Drosophila melanogaster genome. The resulting data were used to retrieve ontologies from Babelomics3. Every functional attribute here described is based on bioinformatically inferred evidence according with functionality based on Drosophila melanogaster which has extensive annotated genome. The Euglossa cordata transcriptome has 49,830 ESTs of which 18,568 related with Apis mellifera genes. Gene Ontology (GO) information was obtained for 5.862 ESTs. The Eulaema nigrita transcriptome has 56,689 ESTs of which 19,354 matched with Apis gene. 5,904 ESTs have GO information with 43% being related with biological processes at level 3. Frieseomellita varia has 50,808 ESTs with 13,795 positively matched against Apis genes. We found 6,070 ESTs being related with biological processes at level 3 and 56% related with molecular function. Melipona quadrifasciata has 54,525 ESTs with 17,521 being related with Apis genes. Interestingly we found GO information for 13,140 ESTs. All species have proportionally the same quantity of ESTs with cellular metabolism, primary metabolism and macromolecules metabolism. They share 4,681 genes based on NCBI non-redundant database.