新西兰兔Nrf2基因的克隆、序列分析及组织表达

为分析Nrf2基因在新西兰兔不同组织的表达差异情况,本试验根据GenBank公布的穴兔Nrf2基因序列设计了特异性引物,从兔肝脏中扩增出Nrf2基因并进行了生物信息学分析,同时采用实时荧光定量PCR技术研究了Nrf2基因在新西兰兔各组织中的表达差异。结果显示,扩增出的新西兰兔Nrf2基因长度为1 758 bp,编码585个氨基酸。序列比对结果显示,新西兰兔与穴兔、人、牛、鼠、斑马鱼等各种属动物Nrf2基因的核苷酸一致性为50.70%~99.83%。进化树分析表明Nrf2在物种间具有较高的保守性。实时荧光定量PCR结果显示,Nrf2基因在新西兰兔的心脏、肝脏、脾脏、肺脏、肾脏和脑中均有表达,且肾脏中表达量最高,脾脏中表达量最低。本试验克隆了新西兰兔Nrf2基因,并研究了其mRNA在兔各组织中的表达情况,为开展兔Nrf2/ARE信号通路的研究奠定了基础。

Cloning and sequence analysis of New Zealand rabbit Nrf2 gene and its expression level in different tissues

To analyze the expression of Nrf2 gene in different tissues of New Zealand rabbits,a pair of primers was designed based on the nucleotide sequences of Oryctolagus cuniculus Nrf2 gene in GenBank.Then Nrf2 gene was amplified from liver of New Zealand rabbit.Using the bioinformation techniques,the gene sequence and the protein structure were analyzed.The expression levels of Nrf2 gene in different tissues were detected by real-time quantitative PCR(qPCR).The results showed that the length of rabbit Nrf2 gene was 1 758 bp and encoded 585 amino acids.The multiple sequence alignment results showed that rabbit Nrf2 gene shared 50.70%-99.83% nucleotide sequence identity with Oryctolagus cuniculus,Homo sapiens,Bos Taurus,Rattus norvegicus,Mus musculus and Danio rerio,respectively.The phylogenetic tree indicated the Nrf2 genes are highly conservative between different species.In addition,the results of qPCR demonstrated that Nrf2 mRNA exists in all detected tissues(heart,liver,spleen,lung,kidney and brain) of New Zealand rabbits.The most abundant expression of Nrf2 was in kidney and the minimal expression was in spleen.Our study lays the foundation for further exploration of rabbit Nrf2/ARE signal pathway.