Ses can reveal an inter-species difference originating from different habitat, feeding
Ses can reveal an inter-species difference originating from different habitat, feeding behavior and other conditions (i.e. L. mucronatus, Figure 6). C. noxius transcriptome analysis was performed by a different sequencing method (pyrosequencing Quizartinib biological activity platform [45]). However, the three conditions used in the library construction allowed comparing theabundance and transcript level differences in each condition and in the same species [45]. At this point, the conditions used for the cDNA library construction are important elements having a repercussion on the ESTs analysis and the interpretation of the differential expression pattern(s). These conditions may also reveal intraspecies (not only inter-species) differences based on the physiological state of the specimen. In recent times, the genome of M. martensii revealed 32,016 protein-coding genes [46]. The authors described a total of 116 neurotoxin genes located in this genome (of which 45 were unknown), consisting of 61 NaTxs, 46 KTxs, 5 ClTxs and 4 CaTx or toxins for ryanodine receptors. In addition, Cao and colleagues [46] confirmed 109 expressed neurotoxin genes in the transcriptome analysis by next generation sequencing (NGS). The advantage and the limitations of the sequencing technology depends on factors such as the sample (venom glands) amount, focus of the study or the cost. But, all sequencing techniques allow to explore different transcriptomes from venomous species. High throughput sequencing or next generation sequencing platforms offer the possibility of generating thousands of sequences that contribute to the study of different conditions and provide a “complete” catalogue of the gene expression (e.g. the 72 toxin-like isogroups from C. noxius represent only 0.4 of the total number of assembled transcripts). In this sense, our low-throughput sequencing is far from a complete catalogue of the gene expression. However, Sanger sequencing in transcriptome is the approachDiego-Garc et al. BMC Genomics 2014, 15:295 http://www.biomedcentral.com/1471-2164/15/Page 11 ofoften used for the screening of the cDNA libraries in the follow conditions: i) limiting sample amount (e.g. one or PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/26240184 two specimens) ii) transcripts sequencing for a future characterization (e.g. cDNA into the vector to future recombinant protein expression) and iii) general catalogues with focus in toxin or venom component transcripts (e.g. selection of the estimated toxin genes by length of the PCR fragments). Our results by Sanger sequencing provided a total of 12 “toxin transcripts” (from 301 clones in the cDNA library) corresponding to 10 of the neurotoxin genes located in the M. martensii genome (or 11 of the expressed neurotoxins by Illumina). Rendon-Anaya et al. [45], identified 72 different toxinlike isogroups from C. noxius analysis by 454 sequencing (e.g. toxins, proteases, antimicrobial peptides) but only 48 PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/26437915 toxin-like isogroups correspond to ion channel specific toxins. Our results of toxin ike transcripts to specific ion channel correspond to 25 of toxin transcriptsobtained by 454 sequencing platform. The number of transcripts and information provided by the transcriptomes by Sanger sequencing is still important for the contribution to the scorpion transcripts.Chlorotoxin-like genes and the first scorpion genome genomic organizationIn the second part of this work, we identified a transcript that encodes a new putative chlorotoxin (Mgib88). This sequence shows a full-length cDNA of 254 bp including the 3′-.
