20.500.12556/DiRROS-21960 Combining short- and long-read transcriptomes for targeted enzyme discovery The discovery of genes that code for a specific enzymatic activity is important in various fields of life science and provides valuable biotechnological tools. Many genes that contribute to the production of secondary metabolites and specialized metabolic pathways are still not identified. Due to the great diversity of metabolic functions found in nature and their rapid evolutionary adaptation, we need precise but high-throughput approaches for a targeted search based on minimal prior knowledge. In this chapter, we describe a transcriptomics pipeline that was used to search for candidate genes coding for a specific enzymatic activity in a nonmodel species. We generated and combined short- and long-read transcriptomic data to obtain reliable full-length transcript sequences along with information on allelic variation, isoform expression, and condition-specific expression. Based on protein domain annotations of coding sequences and transcriptomic data, we selected candidate genes for activity assays. We provide detailed instructions for analysis and quality control steps in our pipeline that can be applied to other biological questions. genes enzymatic activity transcriptomics metabolic pathways allelic variation isoform expression biotechnological tools geni encimska aktivnost transkriptomika presnovne poti alelna variacija izražanje izoforme biotehnološka orodja molekularna biologija molekularna genetika true false false Angleški jezik Slovenski jezik Neznano 2025-04-14 13:41:29 2025-04-14 13:41:29 2025-10-24 03:50:15 0000-00-00 00:00:00 2025 0 0 str. 69-99 0000-00-00 PreprintVObjavi PosredovanoVObjavo NiDoloceno 0000-00-00 0000-00-00 2025-02-04 577 10.1007/978-1-0716-4276-4_4 231281411 231270147 https://link.springer.com/protocol/10.1007/978-1-0716-4276-4_4 1 696bed67-1925-11f0-963a-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=31674 RAZ_Jutersek_Mojca_2025.pdf RAZ_Jutersek_Mojca_2025.pdf 1 A57AE1B9B964888442866FFDB826A5C6 2e4345e8846b674db85e33ba73f6af7bf5e746d36b0253a9e48b5eb8f2ee28cb 86c1f253-1925-11f0-963a-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=31675 Nacionalni inštitut za biologijo Metadata from: Combining short- and long-read transcriptomes for targeted enzyme discovery 0 0 0