鹅脂滴包被蛋白PLIN基因的全长cDNA克隆及组织表达分析

Cloning and Tissue Expressional Pattern of Goose PLIN Gene

脂滴包被蛋白(perilipin, PLIN)是脂滴相关蛋白家族(PAT)中的成员,定位于脂滴表面.该蛋白对脂肪细胞的脂质代谢有重要的调控作用.本研究通过RT-PCR和RACE的方法,从太湖鹅脂肪组织总RNA中扩增出PLIN基因的全长cDNA,测序得到其长度为2 267 bp,其中包括长度为1 584 bp的CDS区,共编码527个氨基酸残基.利用DNAMAN软件将该序列与其他物种的PLIN基因的同源性比较,结果发现鹅PLIN与鸭的同源性最高,达到96.42%,与人的同源性最低,为58.44%.利用MEGA 7.0软件构建基于PLIN基因cDNA序列的系统进化树,显示出PLIN基因的高度保守性.用荧光定量PCR分析其在鹅脂肪、胸肌、腺胃、脑、肾、腿肌、脾、心脏、肺、肌胃、肝实验结果为进一步分析该基因对鹅脂肪性状的作用奠定了基础.

Perilipin(PLIN), a member of the lipid droplet associated protein family(PAT), is located on the lipid droplet surface. Perilipin has important regulatory effects on lipid metabolism in human and mouse adipose cells. In the current study, the c DNA of goose PLIN gene was amplified from the total RNA of adipose tissue by RT-PCR and RACE. The full length c DNA of this gene was 2 267 bp, including a 1 584 bp CDS encoding 527 amino acids. Comparing the homology of the sequence with other species of PLIN gene, the results showed that goose and duck homology was highest at 96%, while goose and human homology was lowest at 58%. The phylogenetic analyses based on PLIN gene CDS showed that the PLIN gene was highly conserved. The detection of expression of PLIN gene in 12 goose tissues of fat, muscle, glandular stomach, brain, kidney, leg muscle, spleen, heart, lung, stomach muscle, liver and small intestine by q RT-PCR showed that PLIN gene was expressed specifically in fat. These results lay a foundation for further analyzing roles of the gene for the fatness characters in geese.