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  

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

作　　者：陈冬金[1] 桑雷[1] 孙世坤[1] 陈敏卿 陈岩锋[1] 王锦祥[1] 谢喜平[1] CHEN Dongjin;SANG Lei;SUN Shikun;CHEN Minqing;CHEN Yanfeng;WANG Jinxiang;XIE Xiping(Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China;Agricultural and Rural Bureau of Longyan,Longyan 364000,China)

机构地区：[1]福建省农业科学院畜牧兽医研究所,福建福州350013 [2]龙岩市农业农村局,福建龙岩364000

出　　处：《畜牧与兽医》2022年第2期15-21,共7页Animal Husbandry & Veterinary Medicine

基　　金：财政部和农业农村部国家现代农业产业技术体系资助(CARS-43-G-5);福建省科技计划项目——省属公益类科研院所基本科研专项(2020R1026004,2021R1026004)。

摘　　要：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2 505 bp(GenBank登录号:MT457404),其中5′非翻译区128 bp, 3′非翻译区223 bp, 2 154 bp的开放阅读框,编码717个氨基酸;序列分析表明,福建白兔与已报道的兔(NM;01082270.1)、人(NM;01750.7)、猴(XM;12638821.1)、猩猩(XM;14344954.2)、小鼠(NM;01301153.1)、大鼠(NM;01033716.1)、猫(XM;11284752.3)、狗(XM;05618083.3)、猪(NM;14067.1)、牛(NM;01030318.3)、绵羊(NM;01009788.1)、山羊(XM;18050015.1)和马(XM;23617937.1)的氨基酸序列同源性分别为99.5%、79.1%、80.4%、79.2%、71.6%、72.6%、77.6%、77.8%、76.8%、76.2%、75.9%、75.9%和78.2%。对13种组织的实时荧光定量PCR分析发现,90日龄福建白兔CAST mRNA表达量以肠组织最高,且极显著高于其他各组织(P<0.01);各组织CAST基因的相对表达量总体趋势为:肠>肺>心、脾、垂体(后三者间表达差异不显著);肾中CAST mRNA表达量显著低于肠、肺、心、脾和垂体(P<0.05),显著高于背肌、下丘脑、胃和肝(P<0.05)。研究结果为进一步开展CAST基因在肉兔肉质改善提供理论参考。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.

关 键 词：福建白兔 钙蛋白酶抑制蛋白 克隆 序列分析 基因表达 

分 类 号：S811.6[农业科学—畜牧学]