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

doi:10.7668/hbnxb.201751529

Abstract

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)

CLC Number:

Q78
S826

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.

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