番鸭GDF9基因克隆、生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2022.03.003

Abstract

Abstract: 【Objective】 The aim of this study was to clone and prediect its structure and function of growth differentiation factor 9 (GDF9) gene of Muscovy duck (Cairina moschata), and to detect its expression differences in Muscovy duck tissues at the nest stage and laying stage.【Method】 The ovary tissue cDNA in Muscovy duck was used as a template, the GDF9 gene was amplified and cloned by rapid-amplification of cDNA ends (RACE).Using biological information software, the similarity of the coding sequence of GDF9 gene was analyzed and evolutionary tree was constructed, and its basic physical and chemical properties and the structural characteristics of the coding amino acid sequence were analyzed.Real-time quantitative PCR was used to detect the expression of GDF9 gene in Muscovy duck of broody period and laying period.【Result】 The total CDS region sequence of GDF9 gene was 1 380 bp, which edcoded 459 amino acids.The similarity of GDF9 gene sequence of Muscovy duck with that of Aythya fuligula, Anas platyrhynchos, Cygnus atratus, Oxyura jamaicensis, Anser cygnodies domesticus, Aptenodytes forsteri, Gallus gallus, Bos taurus, Ovis aries, Sus scrofa and Mus musculus published in GenBank was 98.8%, 97.5%, 96.4%, 95.8%, 94.4%, 90.2%, 87.5%, 64.5%, 64.2%, 63.9% and 60.6%, respectively.The phylogenetic tree showed that Muscovy duck was closely related to Aythya fuligula and Anas platyrhynchos, but far from Mus musculus.The molecular weight of GDF9 protein was 52.81 ku.It was an unstable alkaline hydrophilic membrane protein.It had one signal peptide, 45 phosphorylation sites, 8 O-glycosylation sites, 4 N-glycosylation sites and a transforming growth factor-β (TGF-β) domain.The secondary structure of protein consists of random coil, alpha helix, T-turn and beta sheet composition, accounting for 58.61%, 22.22%, 16.56% and 2.61% respectively.The predicted results of the tertiary structure of GDF9 protein were consistent with the secondary structure.GDF9 protein might interact with BMP15, BMPR1B, BMPR2, TGFBR1, ACVR1B, POF1B, ZP2, ZAR1 and MOS proteins.The results of Real-time quantitative PCR showed that GDF9 gene was expressed in all tissues of Muscovy ducks at nesting and laying stages, the expression of genes in ovary were the highest in two stages, and they were extremely significantly higher than that in other tissues at the same time (P<0.01).Compared with different periods of the same tissue, the expression of GDF9 gene in spleen, heart and kidney in nesting Muscovy duck was significantly or extremely significantly lower than that in laying period (P<0.05 or P<0.01), and the expression in ovary was extremely significantly higher than that in laying period (P<0.01).There was no significant difference in the expression of GDF9 gene in other tissues between the two periods (P>0.05).【Conclusion】 In this study, the GDF9 gene of Muscovy duck was successfully cloned.GDF9 gene was expressed in all tissues of Muscovy duck.GDF9 gene was due to the differentially expressed genes in ovary, heart, kidney and spleen of Muscovy duck during egg laying and nesting.This study provided a theoretical basis for further exploring the function of GDF9 gene in Muscovy duck, and also laid a foundation for studying the biological function and molecular mechanism of GDF9 gene in ovarian development.

Key words: Muscovy ducks; GDF9 gene; ovary; bioinformatics analysis

CLC Number:

SHI Yuzhu, LUO Rutang, CHEN Chao, WU Xu, LI Ang. Cloning, Bioinformatics and Tissue Expression Analysis of GDF9 Genes in Muscovy Duck[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 817-829.

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Cloning, Bioinformatics and Tissue Expression Analysis of GDF9 Genes in Muscovy Duck

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