华北农学报 ›› 2020, Vol. 35 ›› Issue (3): 208-216. doi: 10.7668/hbnxb.20190972

所属专题: 畜牧 生物技术 热点文章

• 畜牧·兽医 • 上一篇    下一篇

牦牛MDH Ⅱ基因克隆表达及脂代谢候选基因关联性分析

陈露露1, 王会1, 王吉坤1, 柴志欣1, 王嘉博1, 陈智华1, 信金伟2, 姬秋梅2, 钟金城1   

  1. 1. 青藏高原动物遗传资源保护与利用四川省、教育部重点实验室, 西南民族大学, 四川 成都 610041;
    2. 省部共建青稞和牦牛种质资源与遗传改良国家重点实验室, 西藏 拉萨 850009
  • 收稿日期:2019-12-12 出版日期:2020-06-28
  • 通讯作者: 钟金城(1963-),男,云南绿春人,教授,博士,主要从事动物遗传资源保护与利用研究;姬秋梅(1965-),女,西藏拉萨人,研究员,博士,主要从事牦牛繁育研究。
  • 作者简介:陈露露(1994-),女,贵州遵义人,硕士,主要从事分子生态学研究。
  • 基金资助:
    国家肉牛牦牛产业技术体系项目(CARS-37);青藏高原生态畜牧业协同创新中心开放基金资助(QZGYXT02);西藏自治区财政专项(XZNKY-2019-C-052)

Cloning and Expression of Yak MDH Ⅱ Gene and Correlation Analysis of Candidate Genes in Lipid Metabolism

CHEN Lulu1, WANG Hui1, WANG Jikun1, CHAI Zhixin1, WANG Jiabo1, CHEN Zhihua1, XIN Jinwei2, JI Qiumei2, ZHONG Jincheng1   

  1. 1. Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Sichuan Province and Ministry of Education, Southwest Minzu University, Chengdu 610041, China;
    2. State Key Laboratory of Barley and Yak Germplasm Resources and Genetic Improvement, Tibet Academy of Agricultural and Animal Husbandry Sciences, Lhasa 850009, China
  • Received:2019-12-12 Published:2020-06-28

摘要: 旨在通过克隆牦牛MDH Ⅱ 基因,探究其组织表达谱及与脂代谢候选基因的相关性,为进一步研究牦牛脂代谢机制提供基础数据。采集0.5,2.5,3.5,7.5岁4个年龄段的12头牦牛心脏、肝脏、肺脏、臀肌和臀脂组织总RNA并反转录成cDNA,实时荧光定量检测MDH Ⅱ 表达量;随机选取12个与脂代谢相关的候选基因,采用实时荧光定量检测MDH Ⅱ 在类乌齐牦牛臀肌、臀脂上的表达量,利用皮尔森系数法计算其与MDH Ⅱ 基因表达量相关性。结果表明:牦牛MDH Ⅱ 基因全长1 196 bp,CDS区长为1 017 bp,编码338个氨基酸,在进化上相对不保守;随年龄增长,MDH Ⅱ 在各组织表达量下降,且在心脏上的表达量高于其他组织;除FABP2VLDLR外的其余10个脂代谢候选基因在牦牛臀脂上的表达量均高于臀肌,MDH Ⅱ 基因在臀肌上的表达量与脂代谢候选基因没有相关性,在臀脂上的表达量与CPT1基因呈显著负相关。本研究成功克隆得到牦牛MDH Ⅱ 基因,其与脂代谢候选基因CPT1显著相关,推测该基因可能参与脂代谢调控,为进一步研究牦牛脂代谢机制提供了理论参考。

关键词: 牦牛, MDH Ⅱ基因, 克隆, 表达, 脂代谢候选基因

Abstract: This study aimed to clone the yak MDH Ⅱ gene, combined with tissue expression analysis and correlation analysis of lipid metabolism candidate genes, which may provide theoretical basis for studying the lipid metabolism mechanism of the yak. 12 yaks (4 age groups:0.5, 2.5, 3.5, 7.5 years old)were selected to extract the heart, liver, lung, gluteal muscle and gluteal fat tissue samples, and total RNA was extracted and reverse transcribed into cDNA. The MDH Ⅱ expression level was detected by RT-qPCR. 12 lipid metabolism candidate genes were randomly selected and their expressions in yak gluteal muscle and gluteal fat was detected by RT-qPCR. Their correlation was analyzed by Pearson coefficient method. The results showed that the yak MDH Ⅱ gene was 1 196 bp in length and the CDS region was 1 017 bp in length, encoding 338 amino acids, which was relatively un-conservative in evolution. With the increase of age, the expression of MDH Ⅱ in each tissue decreased, and the expression in the heart was higher than other tissues. Except FABP2 and VLDLR, the expression of the remaining 10 candidate genes for lipid metabolism on gluteal fat of yak was higher than gluteal muscle. There was no correlation between the expression of MDH Ⅱ gene and candidate genes of lipid metabolism on gluteal muscle. The expression of MDH Ⅱ gene on gluteal lipid was significantly negatively correlated with CPT1 gene. Taken together, we had cloned MDH Ⅱ gene successfully, which was significantly associated with the CPT1 gene. It is speculated that this gene may be involved in the regulation of lipid metabolism, which provides a theoretical reference for further research on the mechanism of lipid metabolism in yak.

Key words: Yak, MDH Ⅱ gene, Clone, Expression, Lipid metabolism candidate genes

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引用本文

陈露露, 王会, 王吉坤, 柴志欣, 王嘉博, 陈智华, 信金伟, 姬秋梅, 钟金城. 牦牛MDH Ⅱ基因克隆表达及脂代谢候选基因关联性分析[J]. 华北农学报, 2020, 35(3): 208-216. doi: 10.7668/hbnxb.20190972.

CHEN Lulu, WANG Hui, WANG Jikun, CHAI Zhixin, WANG Jiabo, CHEN Zhihua, XIN Jinwei, JI Qiumei, ZHONG Jincheng. Cloning and Expression of Yak MDH Ⅱ Gene and Correlation Analysis of Candidate Genes in Lipid Metabolism[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(3): 208-216. doi: 10.7668/hbnxb.20190972.

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