Dissertation > Agricultural Sciences > Crop > Cereal crops > Corn ( maize )

Cloning and Characterization of cDNA of ZmZIP4 Encoding a Zinc Transporter from Maize

Author LiuHaiJun
Tutor HuangYaQun;ChenJingTang
School Agricultural University of Hebei
Course Crop Genetics and Breeding
Keywords Corn Clone Zinc transporter protein OsZIP4 Expression Analysis Subcellular localization The prokaryotic expression
CLC S513
Type Master's thesis
Year 2011
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Zinc is necessary for the metabolism of plant nutrients, its variety of metalloenzymes , transcription factors , as well as other proteins plays a catalytic or building role . Although zinc is essential to plant growth , but an excess of zinc is toxic to plant cells . Therefore , the study of zinc absorption in plants , transportation, storage and distribution has been a concern of many researchers . With the development of molecular genetics, molecular biology and biotechnology , cloning and expression of the gene regulation of plant nutrient physiology research has become one of the important research areas of the post-genomic era plant . This article maize inbred line 178 as the test material , the use of molecular biology methods to clone the zinc transporter gene and its physical and chemical properties , structural characteristics to predict the gene expression characteristics observed in the low - zinc stress and compile product The subcellular localization . Obtained the following results : 1 . Separation rice OsZIP4 ( accession number : AB126089) probe for information homologous cloned into a ZmZIP4 gene , ORF (Open reading frame) of 1161bp, encoding a 386 - amino acid polypeptide chain . The predicted protein has a molecular weight of 38.66 kDa, and the isoelectric point of 7.5 . GenBank number HM048832. 2. Bioinformatics analysis , ZmZIP4 encoding a protein having a ZIP family domain , at the C-terminus contains a signal peptide , having seven transmembrane domains of the encoded protein is a transporter protein . By amino acid sequence comparison, 70% homology found ZmZIP4 with OsZIP4 consistency has reached 73 % , ZmZIP4 OsZIP4 mechanism similar . The semi-quantitative results , ZmZIP4 expression increased when zinc deficiency or copper deficiency or Mn deficiency , instructions may be associated with zinc , copper , manganese transport ZmZIP4 transporter gene . Low zinc stress ZmZIP4 expression peaked three days in the roots , leaves five days to reach the peak , ZmZIP4 zinc stress in roots and leaves of different stress response . 5 Construction of fusion expression the carrier pEGAD - ZmZIP4 gene gun into onion epidermal cells was observed in the cell positioning , the results show that membranes of onion epidermal cells the conversion pEGFP-ZmZIP4 plasmid parts have distinct fluorescence preliminary evidence ZmZIP4 located in the cell membrane.

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