牛ATGL基因的cDNA克隆、序列分析及组织表达研究

cDNA Cloning,Sequence Analysis and Tissue Expression of Bovine ATGL Gene

为了进一步研究牛ATGL基因的结构与功能,揭示该基因的组织特异性表达规律。本研究以秦川牛的脂肪组织为材料,运用同源序列克隆技术结合RT-PCR,对牛的脂肪甘油三酯脂肪酶(ATGL)基因的cDNA进行克隆,并对其进行了生物信息学分析及组织表达研究。结果表明:牛ATGL基因编码区全长为1 461 bp,共编码486个氨基酸;牛ATGL基因与猪同源性最高(87.5%),其次是人(87.3%)、狗(86.8%)、小鼠(84.7%)和大鼠(82.4%);ATGL蛋白含有1个Patatin结构域;牛ATGL基因在脂肪和瘤胃中高丰度表达,在睾丸中没有检测到。ATGL基因在动物进化中比较保守,具有相似的生物学功能,该基因的表达具有组织特异性。

This study was carried out to obtain and analyze sequence of bovine ATGL gene,and to reveal the expression of ATGL mRNA in bovine different tissues.According to the related cDNA sequences of other species ATGL found in the GenBank,the primers of bovine ATGL gene was obtained.The ATGL complete coding sequences in bovine adipose tissue by RT-PCR were obtained and the structure of the bovine ATGL gene was analyzed by bioinformatics methods.The mRNA content was tested in heart,lung,rumen,fat,small intestine,testicle,kidney,pancreas,spleen,liver and muscle tissues of cattle.The results showed that bovine ATGL coding sequence contains 1 461 bp,which encodes 486 amino acids.Bovine ATGL gene exhibited the greatest homology to that of swine（87.5%）,but the lowest to that of mouse（82.4%）.ATGL protein contains a Patatin domain.Bovine ATGL gene was highly expressed in the fat and rumen and was not detected in testicle.The ATGL gene is conservative in the course of animal evolution,and they have approximate biological function in different animals.The expression level of ATGL mRNA is tissue specific.