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Characterization of enhancer trap and gene trap harboring Ac/Ds transposon in transgenic rice
作者姓名:金维正  汪少敏  徐敏  段瑞君  吴平
作者单位:The State Key Laboratory of Plant Physiology and Biochemistry,College of Life Sciences,Zhejiang University Hangzhou 310029,China,The State Key Laboratory of Plant Physiology and Biochemistry,College of Life Sciences,Zhejiang University Hangzhou 310029,China,The State Key Laboratory of Plant Physiology and Biochemistry,College of Life Sciences,Zhejiang University Hangzhou 310029,China,The State Key Laboratory of Plant Physiology and Biochemistry,College of Life Sciences,Zhejiang University Hangzhou 310029,China,The State Key Laboratory of Plant Physiology and Biochemistry,College of Life Sciences,Zhejiang University Hangzhou 310029,China
基金项目:ProjectsupportedbytheHi-TechResearchandDevelopmentProgram(863)ofChina(No.2002AA2Z1003)
摘    要:INTRODUCTIONAfterthewholericegenomewassequencedsuccessfully,moreandmoregenescanbepredictedwithdifferentsoftwaresandannotationsystem.However,thefunctionsofpredictedgeneshavetobefurtheridentifiedbybiologicalanalysis.Oneofthemostpowerfulmethodsassigningfunctiontogeneisthroughinsertionmutagenesiswithtrans-posableelementasDNAsequencetag.Ac/Dstrans-posonsystemhasbecomeaverypopulartoolforgenetaggingandfunctionalgenomicsinvariousplantspecies(Altmannetal.,1995;Chinetal.,1999;Enokietal.,1999;Gre…


Characterization of enhancer trap and gene trap harboringAc/Ds transposon in transgenic rice
JIN Wei-zheng,WANG Shao-min,XU Min,DUAN Rui-jun,WU Ping.Characterization of enhancer trap and gene trap harboring Ac/Ds transposon in transgenic rice[J].Journal of Zhejiang University Science,2004(4).
Authors:JIN Wei-zheng  WANG Shao-min  XU Min  DUAN Rui-jun  WU Ping
Abstract:Insertion mutagenesis has become one of the most popular methods for gene functions analysis. Here we report a two-element Ac/Ds transposon system containing enhancer trap and gene trap for gene tagging in rice. The excision of Ds element was examined by PCR amplification. The excision frequency of Ds element varied from 0% to 40% among 20 F2 populations derived from 11 different Ds parents. Southern blot analysis revealed that more than 70% of excised Ds ele-ments reinserted into rice genome and above 70% of the reinserted Ds elements were located at different positions of the chromosome in rice. The result of histochemical GUS analysis indicated that 28% of enhancer trap and 22% of gene trap tagging plants displayed GUS activity in leaves, roots, flowers or seeds. The GUS positive lines will be useful for iden-tifying gene function in rice.
Keywords:Ac/Ds  Enhancer trap  Excision  Gene trap  Reinsertion  Rice (Oryza sativa L  )  
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