[Aim]  To provide baseline data on the mechanisms underlying silk production in the silkworm, Bombyx mori.  [Methods] This study integrated previous annotations of 15 B. mori genes related to silk production and employed transcriptome analysis to identify those that were significantly differentially expressed. Chromosome mapping, physicochemical property analysis, subcellular localization, and enrichment analysis were then used to elucidate the function of these genes. In addition, interspecies gene family identification and a phylogenetic analysis were performed on the key silk production related genes agtpbp1 and lov of 60 lepidopteran species. [Results]  Silk production related genes NDRG3, PDSS1, beta-Spec, KNCK1, unc-22, BR3, ZMYND8, XNP_1, Bin1 and lov, were significantly differentially expressed in anterior silk gland of B. mori during the dispersal stage compared to 5th instar larva, respectively, with the P-values of 4.523 2× 10^﹣6, 1.586 6×10^﹣4, 5.261 5×10^﹣7, 6.210 8×10^﹣3, 8.113 4×10^﹣10, 2.014 5×10^﹣19, 4.858 2×10^﹣2, 7.767 6×10^﹣23, 3.361 3×10^﹣17, 3.896 7×10^﹣4; beta-Spec, XNP_2, KNCK1, agtpbp1, BR3, XNP_1, Bin1, dpyd and lov, were significantly differentially expressed in middle silk gland of B. mori during the dispersal stage compared to 5th instar larva, respectively, with the P-values of 9.518 4×10^﹣8, 1.984 3×10^﹣3, 4.451 3×10^﹣3, 1.536 7×10^﹣2, 3.185 9×10^﹣8, 1.111 6×10^﹣9, 6.770 7×10^﹣30, 9.297 5×10^﹣3, 4.735 3×10^﹣2; NDRG3, beta-Spec, XNP_2, KNCK1, agtpbp1, unc-22, BR3 and lov, were significantly differentially expressed in posterior silk gland of B. mori during the dispersal stage compared to 5th instar larva, respectively, with the P-values of 8.433 2×10^﹣3, 1.372 5×10^﹣46, 1.059 8×10^﹣10, 6.215 6×10^﹣5, 1.235 0×10^﹣11, 2.491 7×10^﹣3, 8.664 0×10^﹣25, 9.349 1×10^﹣4. These genes directly, or indirectly, influence silk protein synthesis and storage at multiple levels, including transcription, post-transcriptional modification, translation and post-translational modification. The interspecies gene families agtpbp1 and lov contain 34 and 37 genes, respectively, with 26 and 1 positively selected gene in each family. Eight genes, including Bbet001799 in the agtpbp1 family have positively selected sites. [Conclusion]  These results shed light on the function of 15 silk production related genes and the interspecies gene families agtpbp1 and lov.