1 Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China; 2 Fujian Key Laboratory of Animal Genetics and Breeding, Fuzhou 350013, China; 3 College of Animal Science (College of Beeology), Fujian Agriculture and Forestry University, Fuzhou 350002, China
Abstract The 17β-hydroxysteroid dehydrogenase (17β-HSD) eventually oxidizes androstenedione converted from cholesterol to testosterone, which is essential for testosterone synthesis and male reproductive traits. 17β- HSD3 was found to be involved in the pre-embryonic gonadal development of the Mulard duck (Cairina moschata) through the steroid hormone synthesis pathway. In this study, the full-length sequence of 17β-HSD3 gene was obtained by rapid amplification of cDNA ends (RACE) cloning method, sequence characteristics and physicochemical properties of proteins were predicted using bioinformatics analysis, and phylogenetic trees were constructed. The qPCR was utilized to analyze the tissue-specific expression of 17β -HSD3 gene in Mulard duck (brain, heart, liver, lung, muscle, gonad, kidney) and differential expression in the gonads of Mulard ducks, male parent Muscovy and female parent Pekin ducks. The results showed that the full-length sequence of 17β-HSD3 gene was 1 332 bp in Mulard duck, containing a 963 bp of open reading frame, and the length of 5' and 3' non-coding regions were 26 bp and 343 bp, respectively. The encoded protein contained 320 amino acids, which was an unstable protein. Homology and phylogenetic tree phylogenetic analysis showed that the 17β-HSD3 gene in Mulard duck have the closest relationship with swans (Cygnus olor). Tissue-specific expression analysis revealed that the 17β-HSD3 gene was highly significantly more expressed in the gonads of Mulard ducks than in other tissues (P<0.01), suggesting that 17β -HSD3 gene might be involved in gonadal development in Mulard duck. The expression of the 17β -HSD3 gene was significantly higher in Muscovy ducks and Pekin ducks with normal reproductive traits than in Mulard ducks (P<0.01), revealing that 17β - HSD3 gene might be associated with sterility in Mulard ducks. The study provides a theoretical basis for the function of the 17β -HSD3 gene in gonadal development, germ cell differentiation and other reproductive processes in the Mulard duck.
MIAO Zhong-Wei,LI Li,ZHU Zhi-Ming, et al. Cloning, Sequence Features and Expression Analysis of 17β -HSD3, A Key Gene Involved in Gonadal Development of Mulard Ducks (Cairina moschata)[J]. 农业生物技术学报, 2023, 31(4): 801-811.
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