Project description:Many bivalve species produce groups of strong proteinaceous byssal threads to rigidly attach to underwater substrates. Fibres like these have potential applications as biomedical materials due to their unique mechanical characteristics. The byssus and byssal thread producing glands of Pinctada maxima have not yet been characterised. RNA was isolated from P. maxima foot and byssal stem region tissues and sequenced using the Illumina platform. A de novo reference transcriptome comprising 34,281 contiguous sequences was assembled, and tissue replicates were mapped against the reference for quantitative analysis. Tryptic digests of byssal threads were analysed by LC-MS/MS. The resultant peptides were matched to 62 protein sequences derived from our reference transcriptome. Components of the byssus were identified for further characterisation, including a highly expressed perlucin-like foot protein (Pmfp1) and a recently identified protein that we refer to herein as glycine-rich thread (GRT) protein. This work provides principal knowledge on the molecular components of the byssus for P. maxima and the foot ultrastructure involved in the creation of byssal threads. This study advances our knowledge of byssus biosynthesis in non-mytilids, providing a platform for the design of new marine biopolymers.
Project description:The goal of this study was to produce a reference transcriptome for the nickel hyperaccumulator Leucocroton havanensis endemic from Cuba and use this transcriptome as a reference to identify genes responding to the presence or the absence of nickel in both roots and shoots of this species
Project description:Purpose: To demonstrate that gene expression and splicing analysis varies considerably depending on the mapping reference genome. Methods: We mapped and analyzed submitted RNA reads using different tools and reference genomes to evaluate the influence of genome on DEG and alternative splicing tools. Results: We observed that these differences in transcriptome analysis are, in part, due to the presence of single nucleotide polymorphisms between the sequenced individual and each respective reference genome, as well as annotation differences between the reference genomes that exist even between syntenic orthologs. Conclusion: We conclude that even between two closely related genomes of similar quality, using the reference genome that is most closely related to the species being sampled significantly improves transcriptome.
Project description:we applied RNA-seq to detect novel expressed transcripts in 12 tissues of giant pandas, using a transcriptome reconstruction strategy combining reference-based and de novo methods. Then we used mass spectrometry method to identify proteomes of five selected tissues, aiming at validating these novel full-length genes we identified.