广灵驴DGAT2基因克隆、生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2021.02.001

Abstract

Abstract: The study was aimed to clone,bioinformatics of diacylglycerol acyltransferase 2 (DGAT2) gene of Guangling donkey,detected its expression in different tissues,and provided theoretical reference for exploring the mechanism of DGAT2 gene in fat deposition and improving milk fat rate of Guangling donkey.Homologous primers was designed by Primer Premier 3.0 according to the mRNA sequences of DGAT2 gene of Equus caballus (accession No.:XM_023645689.1),Camelus bactrianus (accession No.:XM_010973154.1), Ovis aries(accession No.:XM_027979550.1),and other species published in GenBank.DGAT2 gene sequence of Guangling donkey was amplified with RT-PCR.The encoding sequence of DGAT2 gene was analyzed by bioinformatics methods,and the expression of DGAT2 gene in heart,liver,spleen,lung,kidney,longissimus dorsi muscle,intermuscular fat and subcutaneous fat of Guangling donkey was detected by Real-time quantitative PCR.The results showed that the CDS of Guangling donkey DGAT2 gene consisted of nucleotides of 1 086 bp encoding 361 amino acids.It was submitted to NCBI and the login number was MT993643.The coding sequence of DGAT2 gene showed 99.0%,92.0%,93.5%,92.0%,92.7%,85.3% and 84.1% identity with that of Equus caballus,Bos Taurus,Camelus bactrianus,Sus scrofa,Ovis aries,Homo sapiens and Mus musculus.Phylogenetic tree analysis revealed that Guangling donkey was most closely related to Equus caballus and farthest related to Mus musculus.The DGAT2 protein,with molecular weight of 40.96 ku,fat coefficient of 92.85 and isoelectric point of 9.16,was a stable basic hydrophobic protein with transmembrane region.There were 28 phosphorylation modification sites and 2 glycosylation modification sites in DGAT2 protein,with no signal peptide,it was mainly located in the endoplasmic reticulum.The secondary structure of DGAT2 protein was mainly of 39.89% alpha helix and 36.01% random coil.The DGAT2 gene was expressed in 8 tissues,among which the expression level in subcutaneous fat was significantly higher than that in other tissues (P<0.05),followed by heart,liver and kidney,and the lowest level was in the longissimus dorsi muscle.The experiment results provided a solid theoretical basis for further exploring the role of DGAT2 gene in fat deposition and improving milk quality traits in Guangling donkey.

Key words: DGAT2 gene; Guangling donkey; gene cloning; bioinformatics analysis; tissue expression

CLC Number:

S822

LI Wufeng, GUAN Jiawei, SUN Yutong, QIU Lixia, DU Min. Cloning,Bioinformatics and Tissue Expression Analysis of DGAT2 Gene in Guangling Donkey[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(2): 407-416.

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Cloning,Bioinformatics and Tissue Expression Analysis of DGAT2 Gene in Guangling Donkey

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