Gene Expression Profiling by Quantitative Real-Time PCR (qRT-PCR) A total of 22 and 19 β -hsd genes were collected from the human and zebrafish genome, respectively, which were used to construct HMM models. A set of 19 putative members of the β -hsd genes have been identified from the orange-spotted grouper genome (Table 1). Among the 19 β -hsd genes, a total of 14 were distributed across 10 linkage groups (LGs) in the grouper genome (Figure 1). In addition, we also identified 24, 16, 24, 17, 14, 19, 19, 19, 17, and 27 β -hsd genes in mouse, chicken, xenopus, coelacanth, spotted gar, fugu, tilapia, medaka, stickleback, and common carp, respectively (Table 1). The gene and protein IDs of the β -hsd genes of all species are listed in the attached table (Supplementary Table S1). The exon-intron structure of the grouper β -hsd genes was further characterized, and showed that the number of introns in each β -hsd gene varied from 2–16 (Figure 2B). Hsd3b1 was the gene with the lowest number of introns, containing only two, while hsd17b4 had the greatest amount at 16. In addition, a phylogenetic tree was constructed using the grouper β-HSD protein sequences to determine whether the exon-intron structure is consistent with the phylogenetic tree (Figure 2A).
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