摘要
采用同源克隆、生物信息学方法及荧光定量PCR(Real-time PCR)对缩骨鲫(Carassius auratus sogu var.)肌肉生长抑制素(myostatin,MSTN)基因进行克隆,分析和组织表达状况研究。结果显示:缩骨鲫基因组MSTN基因全长4 737 bp,含有两个内含子和三个外显子;编码区长度为1 128 bp,编码375个氨基酸,包括信号肽(1aa^23aa),TGF-β前肽保守区域(34aa^256aa),TGF-β功能区(281aa^375aa),保守的RXXR蛋白酶酶切位点以及C-端活性区域9个保守的半胱氨酸残基,这与其他物种MSTN相似。此外,缩骨鲫与鲫鱼(Carassius auratus)的MSTN氨基酸序列同源性最高,达到96.1%。以β-actin为内参基因,荧光定量PCR分析表明,MSTN在肌肉中表达量最高;脑、眼、心脏中次之;在肝胰脏、卵巢、肠和肾脏中微量表达。
The myostatin （ MSTN） gene in Carassius a u ratus sogu var. was cloned and analyzed through homology - based cloning, bioinformatics technology and Real - time PCR. The result revealed that the length of MSTN of C. a u ratus sogu var. is 4737bp. It includes 3 exons and 2 introns. The length of coding sequence （ CDS） is 1128bp, the gene code 375 ami-no acids which contain a signal peptide （laa ？23a a ） ,a conserved propeptide domain （ 34aa ？256aa）, a functional do-main （281aa ？375aa）,a RXXR proteolysis site and nine conserved cysteine residues of C - terminal functional domain; As other species, the protein of MSTN have a TGF - p superfamily domain in C. a u ratus sogu var. ; Compared with other species, the homology of MSTN amino acid sequences between C. a u ratus sogu var. and C. a u ratus is closer. Using Real - Time PCR detected MSTN mRNA expression of 9 tissues, the result suggested that the MSTN gene was highly expressed in muscle, second highly expressed in brain, eyes, heart and weakly expressed in ovary, hepatopancreas and kidneys.
出处
《淡水渔业》
CSCD
北大核心
2017年第2期23-29,共7页
Freshwater Fisheries
基金
国家公益性行业(农业)重大专项(201303048)
广东省科技计划项目(2012A020602063
2014A020208145)
