黄羽肉鸡FAT/CD36 cDNA的分子克隆及其发育性表达

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

【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2243bp(GenBank:DQ323177),其中包括1416bp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。

[Objective] This experiment was designed to clone chicken FAT/CD36 complete cDNA sequence and study the ontogenetic 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 from both male and female Yellow-feathered broilers were collected and weighed on d 22, d 29, d 42 and d 56 （n=10）. Fat content of breast and thigh muscles were determined on d 29 and d 56. Real time RT-PCR was conducted to investigate the tissue specific ontogenetic expression pattern of FAT/CD36 mRNA in breast and thigh muscles as well as subcutaneous and visceral fat samples collected from both male and female Yellow-feathered broilers. [Result] Chicken FAT/CD36 complete cDNA sequence of 2 243 bp （GenBank： DQ323177）, which included the 1 416 bp ORF was cloned. The accumulation of subcutaneous and visceral fat levels gradually increased with age. Moreover, the FAT/CD36 mRNA expression levels in subcutaneous and visceral fat of male broilers also increased gradually, the highest of which proved to be in the visceral fat. The FAT/CD36 mRNA expression levels of female broilers were relatively higher during the developmental prophase （d 22 and d 29） whereas a degressive expression pattern was showed during the developmental anaphase （d 42 and d 56）. [Conclusion] Chicken FAT/CD36 complete cDNA sequence was successfully cloned. The expression patterns of muscle and fat in broiler chicken FAT/CD36 mRNA were different with gender.