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To identify candidate β -hsd genes in these thirteen species, we collected the gene and protein names of the β -hsd gene family from the literature (Payne et al., 1997; Dai et al., 2003; Mindnich et al., 2004; Seckl, 2004; Hoffmann and Maser, 2007; Meier et al., 2009; Mindnich and Adamski, 2009; Sreenivasulu and Senthilkumaran, 2009; Rotinen et al., 2010; Kisiela et al., 2012; Saloniemi et al., 2012; Tokarz et al., 2012; Lin et al., 2013; He et al., 2018). Secondly, the amino acid sequences of human and zebrafish β -hsd genes were collected from Ensembl and GenBank by searching for their gene and protein names. Next, specific hidden Markov models (HMMs) of human and zebrafish β-HSDs were developed by hmmbuild command in HMMER (HMMER3.1B2 13 ) and then used as the query sequences to search the protein databases of all the studied species with hmmsearch command in HMMER ( E -value = 0.01). Because all the β -hsd genes belong to the SDR family, except for hsd17b5 , which belongs to the AKR superfamily, there was concern that the HMMs-based search would miss hsd17b5 . Therefore, the amino acid sequence of human HSD17B5 was used as query sequence to search the protein databases of other species by local BLASTP with a cut off E -value of 1e-5 (BLAST 2.2.26 release from NCBI by anonymous FTP 14 ) (Altschul et al., 1997). All the suspected sequences were aligned with the NCBI non-redundant protein sequence database by online BLASTP program to obtain reliable sequences of β -hsd genes. Phylogenetic Analysis. Transcriptome-Based Analysis of Expression Profiling of the β -hsd Genes. Gene Expression Profiling by Quantitative Real-Time PCR (qRT-PCR) Results.
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