链状亚历山大藻铜胺氧化酶基因的克隆和表达分析  

Gene Cloning and Expression Analysis of the Copper-Containing Amine Oxidase from Alexandriumcatenella(Dinoflagellate)

作　　者：商二磊 隋正红[1] 刘源[1] 米萍[1] 刘昊昕 SHANG Er-Lei;SUI Zheng-Hong;LIU Yuan;MI Ping;LIU Hao-Xin(Key Laboratory of Marine Genetics and Breeding(Ocean University of China),Ministry of Education,Qingdao 266003,China)

机构地区：[1]中国海洋大学海洋生物遗传学与育种教育部重点实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2018年第11期79-86,共8页Periodical of Ocean University of China

基　　金：国家自然科学基金项目(41676091);山东省自然科学基金项目(ZR2011DZ002)资助~~

摘　　要：本文以链状亚历山大藻为实验材料,克隆了2个铜胺氧化酶基因,分别命名为cad-1和cad-2。对获取的基因序列进行生物信息学分析,使用实时荧光定量PCR技术分析了cad-1和cad-2在链状亚历山大藻不同生长时期的表达。结果发现,cad-1开放阅读框全长1 023bp,编码340个氨基酸;cad-2开放阅读框全长1 800bp,编码599个氨基酸,含一个内含子1 065bp。2个基因编码的蛋白与颗石藻和抑食金球藻的铜胺氧化酶有较近的亲缘关系。荧光定量PCR分析结果显示,cad-1和cad-2在对数期和衰亡期分别是下调表达和上调表达。依据铜胺氧化酶的功能和链状亚历山大藻衰亡的相关研究,推测铜胺氧化酶可能通过催化多胺产生过量过氧化氢的方式参与链状亚历山大藻赤潮衰亡的过程。Polyamines(PAs)was found widely in plants and are involved in many processes of normal growth and development of all living cells.In plants,PAs participate in the regulation of physiological and developmental processes,and also act as stress messengers responsive to environmental stresses probably.The balance of PAs levels are critical for cells and are regulated by amine oxidases including the flavin-containing polyamine oxidases(PAO)and the copper-containing amine oxidases(CuAO).The diamines are oxidized by CuAO at the primary amine groups,generating corresponding amidoaldehydes,hydrogen peroxide(H 2O 2)and ammoium(NH 3).The H 2O 2 from PAs oxidation is related to programmed cell death(PCD)process.Alexandrium catenella is a representative harmful algal bloom species(HABs),distributed throughtout the world widely.Massive efforts have been made to study on red tide events,however the regulation mechanism of decline phase for red tide caused by A.catenella are still poorly understood.In this study,two genes(cad-1 and cad-2)for copper-containing amine oxidase of A.catenella were cloned by PCR,according to the transcriptomic data.The length of open reading frame(ORF)of gene cad-1 was 1 023 bp,encoding 340 amino acids(Cad-1).The gene cad-2 contained one intron of 1 065 bp and the length of ORF was 1 800 bp encoding 599 amino acids(Cad-2).The results of phylogenetic analysis revealed that Cad-1 and Cad-2 were closely related to CuAOs from both Emiliania huxleyi and Aureococcus anophagef ferens.Quantitative real-time PCR showed that the expression of cad-1 and cad-2 were down-regulated at log phase,and up-regulated at decline phase.Based on the functions of CuAO and the physiological research of the decline phase of A.catenella,it was suggested that CuAO paly certain role in the modulation of decline process of red tide caused by A.catenella.

关 键 词：链状亚历山大藻 衰亡 铜胺氧化酶 实时荧光定量PCR 

分 类 号：S917.3[农业科学—水产科学]