文章摘要
梅高尚,纪德华,李兵,徐燕,陈昌生,谢潮添.坛紫菜核糖体蛋白S15a基因的克隆及表达分析[J].水产学报,2012,36(12):1826~1833
坛紫菜核糖体蛋白S15a基因的克隆及表达分析
Molecular cloning and expression analysis under high temperature stress of ribosomal protein S15a gene from Porphyra haitanensis
投稿时间:2012-04-19  修订日期:2012-09-05
DOI:DOI:10.3724/SP.J.1231.2012.28118
中文关键词: 坛紫菜  ACP技术  核糖体蛋白S15a  克隆  高温胁迫  表达
英文关键词: Porphyra haitanensis  technology of ACP  ribosomal protein S15a  cloning  high temperature stress  expression
基金项目:国家自然科学基金(41176151);十二五“863”计划(2012AA100811);公益性行业(农业)科研专项(200903030);海洋公益性行业科研专项(201105008,201105023);福建省杰出青年基金项目(2010J06016);福建省教育厅新世纪优秀人才项目(JA10186);福建省科技重大专项(2011NZ0001)
作者单位E-mail
梅高尚 集美大学水产学院 meigaoshang@163.com 
纪德华 集美大学水产学院  
李兵 集美大学水产学院  
徐燕 集美大学水产学院  
陈昌生 集美大学水产学院  
谢潮添 集美大学水产学院 ctxie@jmu.edu.cn 
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中文摘要:
      坛紫菜栽培中经常遭遇高温胁迫危害而引起烂菜现象的发生。为研究坛紫菜在高温胁迫条件下的分子应答机制,分离并克隆高温胁迫应答的相关基因,在应用引物退火控制(ACP)技术对耐高温型纯系Z-61叶状体在高温胁迫条件下的差异表达基因进行筛选时,获得了一条在高温胁迫条件下表达水平显著降低的基因片段,并通过5’RACE技术获得了它的全长,命名为Phrps15a(GenBank登录号:JN991055.1)。该基因序列全长676bp,包含一个390bp的开放阅读框编码130个氨基酸的核糖体S15a蛋白(PhRPS15a),蛋白分子式为C664H1066N188O180S7,由3条螺旋,7个片层及8个环状连接组成,与多个物种的RPS15a蛋白具有较高的序列一致性(78~80%)。系统进化分析表明动物、高等植物、绿藻和坛紫菜PhRPS15a蛋白均享有独立的进化分支,但坛紫菜PhRPS15a蛋白与团藻和莱茵衣藻的亲缘关系更近。实时荧光定量PCR分析结果表明Phrps15a基因的表达水平与高温胁迫密切相关,在高温胁迫条件下Phrps15a基因的表达水平极显著下调。
英文摘要:
      Porphyra haitanensis is an economically important marine crop in southern China. A major problem in Porphyra farming is thallus decay in response to high temperature stress. Molecular studies of temperature stress can help to resolve the issue of thallus decay. In this study, to study the molecular mechanism of high temperature tolerance in P. haitanensis, the technology of annealling control primer (ACP) was used to screen the differential expressed genes in gametophytic blades of an F4 high temperature tolerance line Z-61. By the primers combination of dT-RSL and RSL2, one differential expressed gene fragment was cloned. And the full length sequence of this gene fragment was cloned by 5’RACE technology and named as Phrps15a(acceession number JN991055.1). The Phrps15a gene with a 676bp sequences contains an open reading frame of 390bp encoding a PhRPS15a protein of 130 amino acid residues. The PhRPS15a protein which was assembled by 3 helix, 7 sheet and 8 cycle shared high amino acid sequence identity with RPS15 from other organisms (78~80%), and its molecular formula was C664H1066N188O180S7. Phylogenetic analysis showed that the evolution of PhRPS15a of animal, higher plant, green algae and P.haitanensis has unattached evolutional group, and the PhRPS15a protein of P.haitanensis has closer relation with RPS15a proteins from green alga than ones from other organisms. The results of real-time quantitative PCR indiacted that the expressed level of Phrps15a has high relation with high temperature stress. Under high temperature stress, the expression of Phrps15a gene was down-regulated significantly.
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