Meeting Abstract

29.4  Monday, Jan. 4  Transcriptome characterization via 454 sequencing of an oligochaete annelid (Pristina leidyi) used in regeneration research NYBERG, Kevin G.*; BELY, Alexandra E.; University of Maryland, College Park; University of Maryland, College Park kevingnyberg@gmail.com

Recent advances in DNA sequencing technologies have made possible the development of genomic resources in non-traditional model systems. Here, we characterize via 454 pyrosequencing the transcriptome of an oligochaete annelid (Pristina leidyi) that we use to study the evolution of regeneration ability. Batches of worms collected at multiple timepoints during anterior regeneration and posterior regeneration were used to construct a cDNA library consisting of pooled normalized and non-normalized fractions. 454 sequencing of this cDNA library with a Roche GS FLX system and Titanium series reagents resulted in 1.48 million high-quality sequence reads with an average read length of 375 nucleotides. These reads were assembled into 88,358 contigs (average length = 709 nt) with 127,808 singletons (average length = 358 nt). The average contig is of sufficient length to generate an in situ probe without the need for additional sequencing via RACE. Transcript annotation using BLAST and EST2uni and subsequent in situ hybridization has identified a multitude of candidate genes and signaling pathways that may be involved in initiation of regeneration, blastema formation and growth, and morphogenesis in P. leidyi.