2.8 Identification of genes involved in the metabolism pathway in C. striatipennis
The annotated proteins of C. striatipennis were mapped into KEGG’s pathway map to indentify the sequence of all 13 genes or genes family members involved in tricarboxylic acid cycle pathway. And then the required genes from the pathway were deducted. Finally, the extracted protein was verified by KEGG annotation (Kanehisa et al., 2004). Genes and genes families related to porphyrin metabolism, development and metamorphosis were searched using the same method.
2.9 Gene expression analysis
Total RNA of C. striatipennis was respectively extracted from larval, pupal and adult individuals. For each library, three independent replicate RNA samples were prepared. The RNA samples were then sequenced on the illumina platform by biomarker Biotechnology Co., Ltd (Supplementary Table 14). After removing the adapter primers and low-quality reads, the clean reads were obtained for follow-up analysis. The clean reads were mapped to reference genomes by HISAT2 (Kim et al., 2015). The reads mapped on the genome were assembled into complete transcripts by StringTie to evaluate their expression level (Pertea et al., 2015). Differential expression gene set (DEG set) was obtained by differential expression analysis between sample groups using Deseq2 (Love et al., 2014; Trapnell et al., 2012). Then, the functions of differentially expressed genes were annotated based on the Go and KEGG database.