In April 2015, 100 groupers weighing approximately 5 kg were sacrificed for tissue sampling after being anesthetized by MS222. For each fish, the gonad and brain were taken and stored at −80°C for RNA extraction, and a piece of gonadal tissue was fixed in Bouin’s fluid for histological examination. Genome-Wide Identification of β -hsd Genes. 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. Multibet – includes single bets and dicas para o bet365 apostar de futebol accumulator bets.
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