槟榔江水牛OXCT1基因的分子特征及组织表达谱分析

doi:10.16431/j.cnki.1671-7236.2019.06.002

摘要/Abstract

摘要：

本研究对槟榔江水牛3-酮酸辅酶A转移酶1（3-oxoacid CoA-transferase 1，OXCT1）基因的完整CDS区进行了PCR扩增，并对其功能生物信息学和多组织差异表达进行了分析。以普通牛OXCT1基因序列（GenBank登录号：XM_006076397）为参考序列，采用Primer Premier 5.0软件设计引物序列，以提取的槟榔江水牛基因组DNA为模板，PCR扩增获得槟榔江水牛OXCT1基因mRNA序列，扩增产物经测序后利用ORF Finder软件进行开放阅读框识别，获得水牛OXCT1基因CDS区全序列，并对其进行生物信息学分析。结果显示，槟榔江水牛OXCT1基因CDS序列全长1 563 bp，编码520个氨基酸，蛋白分子式为C₂₅₀₉H₄₀₄₁N₆₈₇O₇₄₆S₂₃，分子质量为56.50 ku，理论等电点（pI）为8.69，不稳定系数为27.49，平均疏水性（GRAVY）为-0.097，该蛋白属弱亲水性蛋白。OXCT1蛋白无信号肽和跨膜结构，属于线粒体膜蛋白；包含pcaJ_scoB_fam和AtoD 2个保守结构域，且具有5个功能活性位点。槟榔江水牛与普通牛、藏羚羊等5个物种的OXCT1氨基酸序列同源性≥ 97%。对槟榔江水牛13个组织进行表达谱分析表明，在泌乳期，OXCT1基因在乳腺中表达量最高，在肾脏、垂体、脂肪、大脑、皮肤和肌肉中不表达；在非泌乳期，OXCT1基因在除脂肪以外的其他12个组织中都有表达。OXCT1基因在槟榔江水牛泌乳期乳腺组织中表达量最高，推测其可能参与了水牛的乳脂合成调控事件。本研究结果可为深入了解乳脂合成代谢及其调控机制提供依据。

关键词: 槟榔江水牛; OXCT1基因; 克隆; 生物信息学; 组织表达谱

Abstract:

In this study,the complete CDS of 3-oxoacid CoA-transferase 1 (OXCT1) gene in Binglangjiang buffalo was amplified by PCR method,and the gene functional bioinformatics and tissue differential expression were analyzed.Specific primers were designed by Primer Premier 5.0 according the sequence of Bos taurus OXCT1 gene in GenBank (accession No.:XM_006076397),genomic DNA was extracted from Binglangjiang buffalo as template,the mRNA sequence of buffalo OXCT1 gene was amplified by PCR,the amplification products were sequenced and identified by open reading frame (ORF) with ORF Finder software,and then obtained complete sequence of OXCT1 gene CDS in buffalo which was analyzed by bioinformatics.The results showed that the length of OXCT1 gene CDS was 1 563 bp which encoded 520 amino acids,and the protein molecular formula was C₂₅₀₉H₄₀₄₁N₆₈₇O₇₄₆S₂₃,the molecular weight was 56.50 ku,the theory isoelectric point (pI) was 8.69,the instability coefficient was 27.49,the grand average of hydropathicity was -0.097,and the OXCT1 was the weak hydrophilic protein.The OXCT1 protein had no signal peptide and transmembrane structure,so it belonged to mitochondrial membrane protein.Buffalo OXCT1 contained two conserved domains which named as pcaJ_scoB_fam and AtoD.The homology of OXCT1 amino acid sequences was ≥ 97% between buffalo and other species such as cattle,Tibetan antelope,etc.The expression profile of buffalo OXCT1 gene in 13 tissues was assayed in the stages of lactation and non-lactation.The results showed that the expression level of OXCT1 gene was the highest in mammary gland,but it was not expressed in kidney,pituitary,fat,brain,skin and muscle in lactation.In non-lactating stage,the OXCT1 gene was found to express in all tissues except fat.The OXCT1 gene had the highest expression level of mammary gland in buffalo lactating,which suggested that OXCT1 gene might be associated with the regulation of milk fat synthesis.The results would provide a basis to further understanding about the anabolism and regulation mechanism of milk fat.

Key words: Binglangjiang buffalo; OXCT1 gene; cloning; bioinformatics; tissue expression profile

中图分类号:

Q786

何情, 刘琴, 哈福, 张永云, 苗永旺. 槟榔江水牛OXCT1基因的分子特征及组织表达谱分析[J]. 《中国畜牧兽医》, 2019, 46(6): 1576-1584.

HE Qing, LIU Qin, HA Fu, ZHANG Yongyun, MIAO Yongwang. Molecular Characterization and Tissue Expression Profile of OXCT1 Gene in Binglangjiang Buffalo[J]. , 2019, 46(6): 1576-1584.

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