虹鳟HMGCR基因克隆及组织差异表达分析

Cloning and Tissue Differential Expression Analysis of HMGCR Gene in Oncorhynchus mykiss

[目的]阐述虹鳟β-羟-β-甲基戊二酰辅酶A还原酶(HMGCR)基因的mRNA序列,并比较其在虹鳟不同组织器官中的表达差异。[方法]通过PCR技术扩增虹鳟 HMGCR 基因的mRNA序列,采用荧光定量PCR技术,比较 HMGCR 基因在虹鳟肝脏、肠道、胃、心脏、鳃、脾脏、肾脏、肌肉8种组织器官中的表达差异。[结果]虹鳟 HMGCR 基因的mRNA序列与大西洋鲑的同源性达97%,其蛋白序列与脊椎动物的同源性都在70%以上,表明该基因在物种进化过程中比较保守;该基因在虹鳟肝脏、肠道、胃、心脏、鳃、脾脏、肾脏、肌肉8种组织器官中均有表达;其中,该基因在肝脏和心脏中的表达量较高,而在胃中的表达量最低。[结论]该研究克隆出虹鳟胆固醇合成关键限速酶 HMGCR 的部分mRNA序列,分析了该基因在不同组织中的表达情况,为该酶的深入研究奠定基础。

[Objective]The purpose of this study was to reveal the mRNA sequence of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) and compare the gene expression levels in different tissues of rainbow trout (Oncorhynchus mykiss).[Method]The partial sequence of HMGCR gene was amplified by PCR technology and the gene expression differences among tissues such as liver,intestine,stomach,heart,gill,spleen,kidney and muscle were revealed by fluorescence quantitative PCR technology.[Result]The amino acid sequence shared 97% homology with Salmo salar,and the deduced protein sequence shared more than 70% homology with vertebrate,indicating that the gene was more conservative in the process of species evolution.The gene expression could be detected in the liver,intestines,stomach,heart,gill spleen,kidney and muscle of rainbow trout.The expression level of HMGCR gene in liver and heart was the highest,whereas which was the lowest in the stomach.[Conclusion]We cloned the partial RNA sequence of HMGCR, a key rate-limiting enzyme for cholesterol synthesis in rainbow trout,and analyzed the expression of HMGCR in different tissues,which laid a foundation for further study of the enzyme.