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刺参(Apostichopus japonicus)高温胁迫消减cDNA文库的构建与分析
引用本文:吉成龙,孙国华,杨建敏,宋志乐,刘相全,王卫军.刺参(Apostichopus japonicus)高温胁迫消减cDNA文库的构建与分析[J].海洋与湖沼,2011,42(1):60-66.
作者姓名:吉成龙  孙国华  杨建敏  宋志乐  刘相全  王卫军
作者单位:1. 山东省海洋水产研究所,烟台,264006;上海海洋大学水产与生命学院,上海,201306
2. 山东省海洋水产研究所,烟台,264006
3. 烟台市芝罘区渔业技术推广站,烟台,264001
基金项目:山东省良种工程项目“速生抗病耐高温刺参良种选育”, 2008—2010; 山东省优秀中青年科学家奖励基金(博士基金)项目, 2008BS06004 号; 山东省科技攻关项目“海参新品系选育及构建”, 2008GG10005023 号
摘    要:采用抑制性消减杂交技术(SSH)研究刺参高温胁迫下差异表达的基因.分别以高温实验组为检测组(tester)、常温对照组为驱动组(driver),进行正向抑制性消减杂交;以常温组为tester、高温组为driver,进行反向消减杂交,成功构建了刺参正反双向差异消减文库.从两个文库随机挑选384个白斑克隆进行斑点杂交进一步...

关 键 词:刺参  高温胁迫  抑制性消减杂交  消减文库cDNA
收稿时间:2010/5/10 0:00:00
修稿时间:2010/7/15 0:00:00

CONSTRUCTION AND ANALYSIS OF THE SUBTRACTED cDNA LIBRARY OF APOSTICHOPUS JAPONICUS (SELENKA) UNDER HIGH TEMPERATURE STRESS
JI Cheng-Long,SUN Guo-Hu,YANG Jian-Min,SONG Zhi-Le,LIU Xiang-Quan and WANG Wei-Jun.CONSTRUCTION AND ANALYSIS OF THE SUBTRACTED cDNA LIBRARY OF APOSTICHOPUS JAPONICUS (SELENKA) UNDER HIGH TEMPERATURE STRESS[J].Oceanologia Et Limnologia Sinica,2011,42(1):60-66.
Authors:JI Cheng-Long  SUN Guo-Hu  YANG Jian-Min  SONG Zhi-Le  LIU Xiang-Quan and WANG Wei-Jun
Institution:Shandong Marine Fisheries Research Institute;School of Fishery and Life Science, Shanghai Ocean University;Shandong Marine Fisheries Research Institute;Shandong Marine Fisheries Research Institute;Zhifu Fishery Technical Extension Station;Shandong Marine Fisheries Research Institute;Shandong Marine Fisheries Research Institute
Abstract:Suppression subtractive hybridization (SSH) was used to study the differentially expressed genes of Apostichopus japonicus under high temperature stress. The cDNA of A. japonicus underwent heat stress was used as tester and the same cDNA treated under room temperature was used as driver to construct a forward subtractive library. A reverse subtractive library was generated by using the same cNDA but underwent heat stress was the driver and untreated was tester. 384 positive clones were randomly picked from each library and used for dot blotting. 50 significant differentially expression clones were sequenced and analyzed using BLAST. The results show there are 44 sequences have high identities with the known genes. There are 10 major differentially expressed genes: three up-regulated known genes, two down-regulated known genes, and 5 are unknown genes. The constructed libraries in this study play an important role in studying the heat-resistant genes in A. japonicus and the molecular genetic mechanism of its ability in heat tolerance.
Keywords:Apostichopus japonicus    High temperature stress    Suppression subtractive hybridization    Subtracted cDNA library
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