安氏伪镖水蚤精氨酸激酶cDNA的克隆及镍胁迫下的表达特性  

cDNA Cloning of Arginine Kinase(AK) from Pseudodiaptomus annandalei and the Expression Pattern of AK Under Nickel Stress

作　　者：蒋洁兰[1] 王桂忠[1] 毛明光[1] 李少菁[1] 

机构地区：[1]厦门大学海洋与地球学院,近海海洋环境科学国家重点实验室,福建厦门361102

出　　处：《厦门大学学报（自然科学版）》2013年第4期553-559,共7页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金项目(41276132)

摘　　要：为探讨镍胁迫对桡足类影响的分子机制,以安氏伪镖水蚤(Pseudodiaptomus annandalei)为研究对象,采用cD-NA末端快速扩增技术(RACE)方法获得了安氏伪镖水蚤精氨酸激酶(Arginine kinase,AK)cDNA全序列,全长1 180bp,开放阅读框1 068bp,编码355个氨基酸残基,预测蛋白质分子质量为39.43ku.Blast分析结果显示,安氏伪镖水蚤AKcDNA与罗氏沼虾(Macrobrachium rosenbergii)、多齿新米虾(Neocaridina denticulata)和光滑双脐螺(Bi-omphalaria glabrata)AK基因具有较高的同源性(77%～82%).荧光定量PCR结果显示:低质量浓度镍(10μg/L)对AK转录水平呈现峰值效应,高质量浓度(40和500μg/L)镍则对AK呈现抑制效应,表明AK与安氏伪镖水蚤响应重金属胁迫密切相关.本研究为揭示镍胁迫对桡足类的影响机制提供理论依据.To study the molecular response mechanisms of copepod to nickel stress, the full-length cDNA of arginine kinase gene from the copepod Pseudodiaptomus annandalei was cloned using the strategy of rapid amplification of cDNA ends （RACE）. The ar- ginine kinase cDNA （Pa-AK） was 1 180 bp in length including an open reading frame of 1 068 bp continuously encoded 355 amino acid residues,and the GenBank accession number was JX624126. The molecular weight of the predicted protein was 39.43 ku. Pa-AK showed high homology （77 %-82 % ） with AKs from Macrobrachium rosenbergii, Neocaridina denticulate and Biomphalaria glabra- ta. Further,the transcripts of Pa-AK under nickel stress were examined using the real-time quantitative PCR technique. The results show that：low concentration of nickel （10μg/L） first induced transcripts of Pa-AK, at 12 h, the transcript level started to decreased; high concentrations （40 and 500μg/L） of nickel inhibited transcripts of Pa-AK. The results showed that Pa-AK is closely related to the response to heavy metal stress in Pseudodiaptomus annandalei. The successfully cloning of AK cDNA in Pseudodiaptomus an- nandalei and the study of expression characterization of Pa-AK under nickel stress will facilitate the study of the molecular mecha- nisms of nickel stress on copepod from proteomics aspects.

关 键 词：安氏伪镖水蚤 精氨酸激酶 镍胁迫 表达特性 

分 类 号：Q958.8[生物学—动物学]