SMRT sequencing of the full-length transcriptome of the Sunda pangolin (Manis javanica).

Abstract It is widely known that transcriptional diversity contributes greatly to biological regulation in eukaryotes. With the development of next-generation sequencing (NGS) technologies, several studies on RNA sequencing have considerably improved our understanding of transcriptome complexity. However, obtaining full-length (FL) transcripts remains a considerable challenge because of difficulties in short read-based assembly. In the present study, single-molecule real-time (SMRT) sequencing and NGS were combined to generate the complete and FL transcriptome of Manis javanica. The results provide a comprehensive set of reference transcripts and hence contribute to the improved annotation of the M. javanica genome. We obtained 45,530 high-confidence transcripts from 19,109 genic loci, of which 8014 genes have not yet been annotated within the M. javanica genome. Furthermore, we revealed 8824 long-chain noncoding RNAs (lncRNAs). A total of 30,199 alternative splicing (AS) and 11,184 alternative polyadenylation (APA) events were identified in the sequencing data. The structure and expression level of 59 digestive enzyme genes, including 13 carbohydrase genes, 28 lipase genes and 18 protease genes, were analyzed, which might provide original data for further research on M. javanica. Highlights • It was the first time to report that single-molecule real-time (SMRT) sequencing of Manis javanica. • SMRT sequencing was firstly combined with next-generation sequencing to generate the complete and FL transcriptome of Manis javanica. • We analyzed the expression level and structure of the digestive enzyme genes in the stomach, salivary glands, pancreas, large intestine and liver of Manis javanica. [ABSTRACT FROM AUTHOR]