齐口裂腹鱼PGC-1α基因编码区的克隆及其在肌肉组织中的表达

摘要

Objective: To obtain the peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α) gene se⁃quence of Schizothorax prenanti and study its expression in the red and white muscle tissue of S.prenanti. Meth⁃ods: PCR primers were designed according to Danio rerio PGC-1α gene sequence. Using the total RNA of S.pre⁃nanti muscle tissue and RT-PCR method, we amplified PGC-1α gene sequence, and also analyzed the mRNA ex⁃pression level of PGC-1α gene in red and white muscle of S.prenanti using semi-quantitative RT-PCR. Results:A 2633 bp cDNA fragment(GenBank accession number: JN195738) of S.prenanti PGC-1α gene was obtained. The obtained nucleotide sequence covered an entire ORF of 2631 bp encoding 876 amino acids. Compared with zebraf⁃ish, grass carp and goldfish the homology of amino acid sequences were higher(88%~93%), but lower in some mammals and poultry(49%~50%). The gene expression level of PGC-1α gene in S.prenanti red muscle basal me⁃tabolism statu was significantly higher than that of white muscle(P<0.05), and PGC-1α was induced in S.prenanti red and white muscles under fasting condition. Conclusion: We firstly obtained the PGC-1α gene sequence of S. prenanti, and it's expression level in red muscle was significantly higher than that in white muscle, and PGC-1αwas induced in red and white muscles by fasting. Our results will provide important theoretical basis for the role of PGC-1α on the muscle tissue of S.prenanti.%　　目的：获得齐口裂腹鱼过氧化物酶体增殖物激活受体γ辅助活化因子1α(PGC-1α)cDNA序列,探讨其在齐口裂腹鱼肌肉组织中的表达规律。方法：根据斑马鱼基因PGC-1α序列设计引物,提取齐口裂腹鱼肌肉组织总RNA,经RT-PCR扩增PGC-1α基因序列；利用半定量RT-PCR分析齐口裂腹鱼PGC-1α基因在红肌和白肌中的mRNA表达特性。结果：获得齐口裂腹鱼PGC-1α基因序列2633 bp,GenBank登陆号为JN195738,其中ORF为2631 bp,编码876个氨基酸残基,与同鲤科的斑马鱼、草鱼和金鱼的同源性较高,为88%~93%,但与哺乳动物和禽类如人、小鼠、大鼠、猪、牛和鸡的同源性较低,为49%~50%。在基础状态下,PGC-1α在齐口裂腹鱼红肌中的表达显著高于白肌,禁食可显著诱导PGC-1α在红肌和白肌中的表达水平。结论：首次克隆得到齐口裂腹鱼PGC-1α基因序列,其在红肌中的表达显著高于白肌中,禁食可显著诱导其在红肌和白肌中的表达,为研究PGC-1α在齐口裂腹鱼肌肉中的作用提供了理论依据。