黄羽肉鸡FAT/CD36 cDNA的分子克隆及其发育性表达

 【目的】克隆鸡FAT/CD36 cDNA的全长序列并探讨FAT/CD36 mRNA的发育性表达。【方法】采用cDNA末端快速扩增法（RACE）克隆鸡FAT/CD36 cDNA的全长序列。选用22、29、42和56日龄的黄羽肉鸡公鸡和母鸡各10羽，分离皮下脂肪和腹脂并称重，同时测定22和56日龄时胸肌和腿肌的脂肪含量。分别采集胸肌、腿肌、皮下脂肪和腹脂样品，采用实时荧光定量RT-PCR方法检测了FAT/CD36 mRNA的表达。【结果】鸡FAT/CD36 cDNA的序列全长为2 243 bp（GenBank：DQ323177），其中包括1 416 bp开放阅读框（ORF）。黄羽公鸡皮下脂肪和腹脂的沉积量随日龄的增加逐渐升高，腿肌和腹脂FAT/CD36 mRNA 的表达水平也逐渐升高，其中腹脂的表达水平在所检测的各组织中最高；母鸡FAT/CD36 mRNA 的表达水平在生长早期（22和29日龄）较高，但在后期（42和56日龄）反而有下降的趋势。【结论】本研究成功地克隆了鸡FAT/CD36 cDNA的全长序列，黄羽肉鸡肌肉和脂肪组织FAT/CD36的表达存在性别差异。

Molecular Cloning and Ontogenetic Expression of Fatty Acid Translocase cDNA in Yellow-Feathered Broiler

Abstract: 【Objective】To clone chicken FAT/CD36 complete cDNA sequence and study the ontogenesis expression pattern of FAT/CD36 mRNA;【Method】Chicken FAT/CD36 complete cDNA was cloned by 5’- and 3’-RACE. Subcutaneous fat and visceral fat samples were collected and weighed from both male and female Yellow-feathered broilers on d22, d29, d42 and d56 (n=10). Fat content of breast muscle and thigh muscle were determined on d29 and d56. Real time RT-PCR was conducted to investigate the tissue specific ontogenesis expression pattern of FAT/CD36 mRNA in breast muscle, thigh muscle, subcutaneous fat and visceral fat samples collected from both male and female Yellow-feathered broilers;【Result】We have cloned chicken FAT/CD36 complete cDNA sequence of 2243bp(GenBank: DQ323177), which including the 1416bp ORF. The accumulation of subcutaneous fat and visceral fat gradually increased associated with age and the FAT/CD36 mRNA expression levels in subcutaneous fat and visceral fat of male broilers also increased gradually, the highest expression level proved to be in the visceral fat. The FAT/CD36 mRNA expression levels of female broilers were relatively high during the development prophase (d22 and d29), whereas a degressive expression pattern was showed during the development anaphase (d42 and d56);【Conclusion】We have successfully cloned chicken FAT/CD36 complete cDNA sequence. The expression pattern of FAT/CD36 mRNA in muscle and fat were different with gender.