Cloning and Expression of Goat (Capra hircus) CPT1B Gene and Its Correlation with Intramuscular Fat Content
YU Yu-Yang1, LIN Ya-Qiu1, LIANG Ji-Jun1, WANG Yong1, 2, *, ZHU Jiang-Jiang2, 3, *
1 College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China; 2 Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization Key Laboratory of Sichuan Province, Chengdu 610041, China; 3 Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization (Southwest Minzu University), Ministry of Education, Chengdu 610041, China;
Abstract：Intramuscular fat content is a key indicator of animal breeding. It is an important task to study the regulation mechanism of lipid metabolism by molecular biology. The present study aims to clone the carnitine palmityl transferase 1 B gene (CPT1B) and determine its expression in various tissues and intramuscular precursor adipose cells of goats (Capra hircus), which will lay the foundation for further study of CPT1B gene in regulating goat fatty acid metabolism. Seven healthy Jianzhou Big-eared goats aged 1 year old were selected, and the tissue samples of heart, liver, spleen, lung, kidney, back longissimus dorsi, biceps femoris, triceps, subcutaneous fat and abdominal fat were quickly collected and stored in liquid nitrogen. The intramuscular fat contents of back longissimus dorsi, biceps femoris, triceps were detected by Soxhlet extraction method. The total RNA of the above-mentioned tissue samples was extracted using Trizol reagent and the sequence of CPT1B was cloned by RT-PCR following by the sequence bioinformatics analysis. The expression of CPT1B in various muscle tissues were determined by qRT-PCR method, which were then used for correlation analysis with the intramuscular fat content. The results showed that the goat sequence was 2 619 bp (GenBank No. MH340532), including 2 313 bp CDS, 52 bp 5'UTR and 254 bp 3' UTR, encoding 771 amino acid residues. Tissue quantification results showed that CPT1B was expressed higher in heart, longissimus dorsi, biceps femoris and tricep, and expressed lower in liver, spleen, lung, kidney, subcutaneous fat and abdominal fat of Jianzhou Big-eared goat. The qRT-PCR results of intramuscular precursor adipose cells showed that the expression of CPT1B during differentiation of preadipocytes was significantly higher than that before and after differentiation (P<0.01). The expression of CPT1B gene was significantly positively correlated with the intramuscular fat content in all the 3 muscle tissues (P<0.05). In summary, CPT1B might have a regulatory role in goat intramuscular fat deposition, which also lays a foundation for the application of CPT1B gene in meat goat breeding.
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