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Functional Analysis of OsSEC27p and Cloning, Subcellular Localization of OsYSL1

Author: MaFeng
Tutor: YinLiPing
School: Capital Normal University
Course: Cell Biology
Keywords: Rice Iron Deficiency Vesicular transport Subcellular localization Non -invasive ion scan technology FM4-64
CLC: Q943.2
Type: Master's thesis
Year: 2007
Downloads: 207
Quote: 0
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Abstract


The papers for my lab early gene chip study results of two raised the Iron Deficiency rice root to express the gene OsSec27p and OsYSL1 respectively cloning , subcellular localization and functional analysis . Iron deficiency 5 days to process rice root cDNA microarray and realtime-PCR analysis detected high expression OsSec27P . Than the chip in the original EST sequences found the the a slight homologous therefore the genes with yeast Sec27P named OsSec27P . The full-length eDNA OsSec27p 's has been cloned and submitted to the GenBank accession and accession number DQ434847 . Gfp reporter gene and molecular probe FM4-64 , its subcellular localization , found The pCAMBIA1302-OsSEC27p :: GFP is located in the plasma membrane and endomembrane system small vesicles . Subsequently , using Agrobacterium mediated into tobacco and T 1 and T 2 generation of plant molecular detection and physiological indicators detected . Application of non-invasive ion scan technology transgenic tobacco radicle H < / sup> flow detection , found OsSec27p gene and outgoing proton secretion . On the other hand , cloning OsYSL1 and build plants the expression vectors pCAMBIA1302-OsYSL1-gfp and expression the the carrier pGPTV - AGPp - BAR- OsYSL1 . Recombinant plasmid pCAMBIA1302-OsYSL1-gfp into onion epidermal cells and BY-2 tobacco suspension cells , and cellular localization studies , results show OsYSL1 located in the cytoplasmic membrane . At the same time , restructuring the plasmid pGPTV - AGPp -BAR - OsYSL1 into transgenic rice calli experiment . The found the corn ZmYS1 have 43.43% homology sequence analysis of OsYSL1 . Therefore , we concluded that it might transporter protein of a high-speed rail carrier and ZmYS1 .

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CLC: > Biological Sciences > Botany > Plant Cell Genetics > Plant Genetic Engineering
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