• Pigment distribution were different between yellow- and white-cocoon silkworm MSGs. • DEGs, DEPs and Co-DEGs-DEPs were identified from yellow-cocoon silkworm strains. • Key factors such as CBP and Cameo2 in BmCSTS were verified in this study. • A predominant ribosome-biogenesis regulation network functions during cocoon color formation. The human macular carotenoid transporter was firstly discovered using silkworm yellow-cocoon strains, known as Bombyx mori carotenoid selective transport system (BmCSTS). However, molecular mechanisms underlying the gene/protein expression and regulations in vivo remain elusive. Here, 620 differentially expressed genes (DEGs), 1965 differentially expressed proteins (DEPs), and 67 co-genes (namely Co-DEGs-DEPs) from yellow-cocoon silkworm middle silk glands (MSGs) were identified by RNA-Seq based transcriptome and SWATH-based proteomic analysis. Notably, previously identified CBP-alike key factors were further demonstrated significantly up-regulated in all six B. mori yellow-cocoon strains (NB, NB × 306, 306 × NB, NB × 798, 798 × NB, and XH × NB) used in this study. The GO and KEGG analysis results clustered most DEGs and DEPs into multiple biological processes including both cellular components and molecular functions, especially protein synthesis and energy-metabolic pathways. The protein–protein interaction (PPI) network analysis results further revealed a predominant ribosome-biogenesis regulation working system during silkworm yellow-cocoon color formation. Additionally, high-efficient energy production and metabolism pathways (i.e. oxidative phosphorylation and citrate cycle) were also discovered, which might facilitate key gene/protein expressions and functions in BmCSTS, such as CBP and Cameo2.