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Exploring Toxin Evolution: Venom Protein Transcript Sequencing and Transcriptome-Guided High-Throughput Proteomics.

Methods Mol Biol. 2020; 
Modahl CM1,2, Durban J3, Mackessy SP4.
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DNA Sequencing … additive preparation. 9. Quick Clean 5 M Miniprep kit (<b>Genscript</b> ® ) or similar plasmid<br> purification kit. 2.3 Next-Generation Sequencing (NGS) Transcriptomics. 2.3.1 NGS<br> Library Preparation and Data Generation. 1. TruSeq RNA … Get A Quote

摘要

Studying animal toxin evolution requires sequences of these proteins and peptides, and transcript sequences allow for the construction of cladograms and evaluation of selection pressures from nonsynonymous and synonymous nucleotide mutation ratios. In addition, these translated sequences can be useful as custom databases for peptide identifications within venoms and for better proteomic quantification. Obtaining these transcripts is achieved by sequencing cDNA originating from venom gland tissue or venom. This chapter provides the methodology for (1) targeted sequencing of transcripts from a single venom protein family (RNA isolation and 3'RACE [rapid amplification of cDNA ends]), (2) generation of a venom glan... More

关键词

3′RACE; Bioinformatics; Next-generation sequencing; Proteomics; Toxin evolution; Transcriptomics; Venomics