山羊<i>CPT1A</i>基因的克隆表达及肌内脂肪含量的相关性分析

doi:10.7668/hbnxb.201751529

摘要/Abstract

摘要： 旨在获得山羊肉毒碱棕榈酰基转移酶Ⅰ肝脏亚型CPT1A（Carnitine palmitoyltransferase 1A，CPT1A）基因序列，分析其生物学特征，阐明其组织及细胞分化时序表达规律，同时揭示CPT1A基因在背最长肌、股二头肌、臂三头肌中与肌内脂肪含量（IMF）的关系。选取7只1周岁健康简州大耳羊为试验动物，屠宰后迅速采集心、肝、脾、肺、肾、背最长肌、股二头肌、臂三头肌、皮下脂肪和腹间脂肪组织样品，利用TRIzol法提取总RNA。采用RT-PCR法克隆山羊CPT1A基因，进行生物信息学分析。利用实时荧光定量PCR技术检测CPT1A基因在不同组织中的表达水平以及CPT1A在山羊前体脂肪细胞分化过程中的时序表达。使用皮尔逊相关系数进行CPT1A基因表达量与IMF含量相关性分析。通过克隆得到CPT1A基因序列（MH345735）为2 380 bp，包含CDS全长2 319 bp，5'UTR 38 bp和3'UTR 23 bp，编码773个氨基酸。组织表达结果显示，CPT1A在简州大耳羊肝脏和肾脏中的表达水平极显著高于其他组织（P<0.01）。细胞时序表达定量结果显示，CPT1A在诱导分化0~5 d逐渐升高，在第5天表达量最高（P<0.01），5~9 d逐渐下降。CPTA基因表达量与IMF含量相关性分析结果显示，CPT1A基因表达量与各肌肉组织肌内脂肪含量均显著正相关。结果表明，CPT1A可能在肌内脂肪沉积过程中具有重要调控作用，为进一步研究CPT1A调控山羊脂质代谢的机理研究提供参考。

关键词: 山羊, CPT1A基因, 克隆, 组织表达, 肌内脂肪

Abstract: The aim of this study was to obtain the gene sequence of goat carnitine palmitoyl transferase I liver subtype CPT1A (Carnitine palmitoyltransferase 1A, CPT1A), analyze its biological characteristics, and elucidate its tissue expression and cell differentiation temporal expression. Besides, the relationships between the expression of CPT1A gene and intramuscular fat content (IMF) in the longissimus dorsi, biceps femoris and triceps were revealed. Seven one-year-old healthy Jianzhou Big-Eared goats were used as the test animal. After slaughter on an empty stomach, the heart, liver, spleen, lung, kidney, longissimus dorsi, biceps femoris, triceps, subcutaneous fat and abdominal fat tissue were collected rapidly. Total RNA was extracted using the TRIzol method. The goat CPT1A gene was cloned by RT-PCR for biological analysis. Real-time PCR was used to detect the expression level of CPT1A gene in different tissues and the temporal expression of CPT1A in the differentiation of goat preadipocytes. The correlation between CPT1A gene expression and IMF content was analyzed by Pearson correlation coefficient. The cloned C PT1A sequence (MH345735) was 2 380 bp, containing the full-length CDS 2 319 bp, 5'UTR 38 bp and 3'UTR 23 bp, encoding 773 amino acids. The results of tissue expression showed that the expression level of CPT1A in the liver and kidney of Jianzhou Big-Eared goat was significantly higher than in other tissues (P<0.01). Quantitative results of cell temporal expression showed that the expression level of CPT1A gradually increased in 0-5 d, reaching to the highest at 5 d (P<0.01), and then decreased gradually in 5-9 d. Correlation analysis between CPT1A gene expression and IMF content showed that CPT1A gene expression was significantly positively correlated with intramuscular fat content in each muscle tissue. These results indicated that CPT1A might play an important regulatory role in intramuscular fat deposition, and facilitate further study on the mechanism of CPT1A regulation in goat lipid metabolism.

Key words: Goat, CPT1A gene, Clone, Organizational expression, Intramuscular fat(IMF)

中图分类号:

Q78
S826

梁计峻, 林亚秋, 俞雨阳, 王永, 朱江江. 山羊CPT1A基因的克隆表达及肌内脂肪含量的相关性分析[J]. 华北农学报, 2019, 34(5): 231-238. doi: 10.7668/hbnxb.201751529.

LIANG Jijun, LIN Yaqiu, YU Yuyang, WANG Yong, ZHU Jiangjiang. Cloning and Expression of Goat CPT1A Gene and Its Correlation with Intramuscular Fat Content[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(5): 231-238. doi: 10.7668/hbnxb.201751529.

http://www.hbnxb.net/CN/Y2019/V34/I5/231

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Cloning and Expression of Goat CPT1A Gene and Its Correlation with Intramuscular Fat Content

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