高原花斑裸鲤Cu/Zn-SOD的克隆鉴定表达及其对Cu^(2+)胁迫的应答

Cloning,Identification and Expression of Cu/Zn-SOD and Its Responses to Cu^(2+) Stress in Plateau Gymnocypris eckloni

目的对高原花斑裸鲤(Gymnocypris eckloni)铜锌-过氧化物岐化酶(Cu/Zn-Superoxide Dismutase,Cu/Zn-SOD)基因的克隆鉴定表达及对其Cu^(2+)胁迫的应答分析。方法采用RT-PCR和RACE技术获得G.eckloni Cu/Zn-SOD全长c DNA序列,分析生物信息学特征和在不同组织的表达分布。在Cu^(2+)的胁迫下,通过实时定量PCR检测花斑裸鲤肝、肾、脑、鳃组织中Cu/Zn-SOD的表达变化,探究其是否能够作为响应重金属污染胁迫的一种生物标记物。结果Cu/Zn-SOD的cDNA序列全长785 bp,其中3'-UTR为277 bp,5'-UTR为43 bp,编码区为465 bp,可编码154个氨基酸(Gen Bank登录号为:KX826080)。在Cu^(2+)胁迫下,花斑裸鲤肝、肾、脑、鳃组织中的Ge Cu/Zn-SOD在mRNA水平相对表达量呈现出一定的规律性。结论成功实现高原花斑裸鲤Cu/Zn-SOD的克隆鉴定表达,初步得到了花斑裸鲤Cu/Zn-SOD对Cu^(2+)胁迫的应答模式。

Objective To investigate the cloning,identification and expression of Cu/Zn-SOD and analysis its responses to Cu2＋ stress in plateau Gymnocypr/s eckloni.nethods The full-length cDNA of Cu/Zn-SOD in G.eckloni was cloned by means of RT-PCR and RACE method, and the molecular bioinformatics characterization and tissue distribution were analyzed.Under the stress of Cu2＋ ,the relative expression level of Cu/Zn-SOD in G.eckloni in liv- er,gill,brain and kidney tissues were detected through real-time quantitative PCR,to explore whether it can be used as a biomarker to respond to heavy metal pollution stress.Results The results showed that the full-length cDNA sequence of Cu/Zn-SOD was 785 bp, containing a 277 bp 3＇-untranslated region （UTR）, a 43bp 5＇-UTR and a465 bp open reading frame （ORF）, encoding 154 amino acids（ GenBankaccession number： KX826080）.The respon- ding model of Cu/Zn-SOD in G. eckloni under the stress of Cu2＋ was analyzed, in which the relative expression lev- els of C, eCu/Zn-SOD in G. eckloni in liver, gill,brain and kidney showed some regular trends of change. Conclusions The successful acquisition of cloning,identification and expression of Cu/Zn-SOD and its responses to Cu2~ stress in plateau G.eckloni were obtained preliminarily.