福建白兔钙蛋白酶抑制蛋白基因的克隆及其表达差异研究

为了解福建白兔钙蛋白酶抑制蛋白(calpastatin,CAST)基因分子基础,应用RT-PCR和克隆测序从肌肉组织总RNA中克隆出福建白兔CAST基因的cDNA序列并进行分析.通过荧光定量PCR测定福建白兔不同组织CAST基因mRNA表达谱.结果 显示:该基因序列长度为2505 bp(GenBank登录号:MT457...

Cloning and differential expression of the calpastatin gene in Fujian white rabbit

In order to understand the molecular basis of the calpastatin(CAST) gene in Fujian white rabbits, the cDNA sequence of the gene was cloned from the total RNA of the muscle tissues of the rabbits by RT-PCR and sequencing. The mRNA expression profiles of the CAST gene in different tissues of the Fujian white rabbits were determined by fluorescence quantitative PCR. The results showed that the full length of CAST cDNA(GenBank accession number: MT457404) was 2 505 bp encoding a peptide of 717 amino acid. The amino acid homology of the Fujian white rabbit CAST with reported rabbit(NM;01082270.1), human(NM;01750.7), monkey(XM;12638821.1), orangutan(XM;14344954.2), mouse(NM;01301153.1), rat(NM;01033716.1), cat(XM;11284752.3), dog(XM;05618083.3), pig(NM;14067.1), cattle NM;, sheep(NM;01009788.1), goat(XM;18050015.1) and horse(XM;23617937.1) were 99.5%, 79.1%, 80.4%, 79.2%, 71.6%, 72.6%, 77.6%, 77.8%, 76.8%, 76.2%, 75.9%, 75.9% and 78.2%, respectively. The real-time fluorescence quantitative PCR analysis of the 13 kinds of tissues showed that the mRNA expression of CAST in the intestinal tissues was the highest in the 90-day-old Fujian white rabbits, which was very significantly higher than that in the other tissues(P<0.01). The general trend of the relative expression of the CAST gene in the tissues was as follows: intestine>lung>heart, spleen and pituitary(no significant difference among the last three). The expression in kidney was very lower than that in intestine, lung, heart, spleen and pituitary, and was higher than that in the dorsal muscle, hypothalamus, stomach and liver(P<0.05). These results provided a theoretical basis and reference for further research on the CAST gene in improving the meat quality of rabbits.