广灵驴ADSL基因克隆、序列分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2021.08.001

Abstract

Abstract: The purpose of this study was to clone and analyze of adenylosuccinatelyase(ADSL) gene in Guangling donkey by bioinformatics, detect its expression in different tissues, and provide theoretical reference for exploring the action mechanism of ADSL gene in inosine monophosphate and flavor formation of Guangling donkey.Homologous primers were designed by Primer Premier 3.0 according to the mRNA sequences of ADSL gene in Equus caballus (accession No.:XM_001917207.5), Bos taurus (accession No.:NM_001102377.2) and Sus scrofa (accession No.:GU249574.1), and other species published in GenBank.The CDS sequence of ADSL gene in Guangling donkey was amplified with RT-PCR and cloned.The encoding sequence of ADSL gene was analyzed, and the expression of ADSL gene in heart, liver, spleen, lungs, kidney, longissimus dorsi muscle and subcutaneous fat of Guangling donkey were detected by Real-time quantitative PCR.The CDS of ADSL gene in Guangling donkey consisted of nucleotides of 1 473 bp, encoding 490 amino acids.It was submitted to NCBI, and obtain the accession No.:MW037837.The nucleotide sequence of ADSL gene showed 99.5%, 90.8%, 92.3%, 90.4%, 90.7%, 90.5% and 86.0% identity with that of Equus caballus, Bos taurus, Camelus bactrianus, Sus scrofa, Ovis aries, Homo sapiens and Mus musculus, respectively.Phylogenetic tree results revealed that Guangling donkey was the most closely related to Equus caballus and the farthest related to Mus musculus.The ADSL protein, with molecular weight of 55.44 ku, isoelectric point of 6.52, and grand average of hydrophobicity of -0.243, was an unstable acidic hydrophilic protein.There were 41 phosphorylation modification sites, 6 glycosylation modification sites, and a coiled helix in ADSL protein, with no signal peptide and transmembrane structure, it was mainly located in the cytoplasmic.The secondary structure of ADSL protein was mainly of 68.98% alpha helix.ADSL gene was expressed in 6 tissues, among which the expression in lung was the highest, which was significantly higher than that in other tissues(P<0.05), followed by heart and liver, and the lowest in spleen, kidney and longissimus dorsi muscle.The experiment results provided a solid theoretical basis for further exploring the role of ADSL gene in the molecular mechanism of inosine monophosphate synthesis and flavor formation in Guangling donkey.

Key words: Guangling donkey; ADSL gene; cloning; bioinformatics analysis; tissue expression

CLC Number:

GUAN Jiawei, SUN Yutong, QIU Lixia, LI Wufeng, DU Min. Cloning,Sequence Analysis and Tissue Expression of ADSL Gene in Guangling Donkey[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2685-2694.

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Cloning,Sequence Analysis and Tissue Expression of ADSL Gene in Guangling Donkey

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