Dissertation > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering)

Proteome and Transcriptome Maps Analysis of Transformed Yeast with MxIRT1 or OsNramp1

Author ZhangWenJuan
Tutor YinLiPing
School Capital Normal University
Course Cell Biology
Keywords MxIRT1 OsNramp1 Two-dimensional electrophoresis of proteins Peptide mass fingerprinting Gene chip Vesicular transport
Type Master's thesis
Year 2007
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MxIRT1, and OsNramp1 is a metal transporter protein was cloned the from small Jinhaitang and rice root. Heterologous functional complementation experiments show that the two genes are complementary the yeast two mutant fet3fet4 in the absorption of iron. In this study, proteomic and transcriptional level of the group of two genes and their changes caused. The first part of the study dimensional gel electrophoresis and peptide mass fingerprinting technology IRT strains turn over iron and iron deficiency conditions, respectively, under the conditions of normal iron processing turn Nramp and turn empty vector strain normal iron processing and treatment of iron deficiency conditions The following were transfected with the empty carrier, IRT and Nramp yeast strains proteome analysis. Protein two-dimensional electrophoresis for each treatment under the conditions of yeast strains can be isolated spots around 1500. Turn the IRT strains retrieval get over iron and iron deficiency conditions 8 differentially expressed proteins with the cytoskeleton, chaperone, synthesis of amino acids, glycerol synthesis related; the Nramp turn normal iron processing conditions and transfected with empty vector strain retrieved nine differentially expressed proteins were associated with the molecular chaperone, glucose metabolism, the synthesis of spermidine, anti-oxidation; normal iron processing and treatment of iron deficiency conditions were transfected with empty vector, the IRT and Nramp yeast strains retrieved 11 differentially expressed proteins, in addition to a protein of unknown function, the other 10 difference and glucose metabolism, protein biosynthesis, cell anti-poison, vesicular transport and signal transduction proteins. The second part of the study using gene chip technology on the normal iron processing conditions were transfected with empty carrier, IRT and Nramp yeast strains transcription of this group. Yeast extract total RNA, reverse transcription of a biotin label, 10928 yeast cDNA microarray hybridization. Turn IRT strains transfected with empty vector strain regulated genes relative to 1200, down-regulated genes 1140; genetically Nramp strains transfected with empty vector strain regulated genes relative to 290, down-regulated genes 108 months. On the the Nramp turn strains the difference is more significant 27 up-regulated genes and 45 down-regulated gene expression analysis found upregulated gene polymorphism with the zinc finger structure associated with RNA-binding regulator or inhibit the child and hypertonic induced expression The genes downregulated gene and vesicular transport, cell communication, cell cycle, and protein synthesis. Speculated that in genetically modified yeast, the IRT or Nramp transferred, single pleiotropic role more obvious. Significant changes in addition to the effect of DNA replication, RNA transcription, protein synthesis, oxidation respiratory role, transcription factors, molecular chaperones there are specific molecular structure change, vesicular transport-related genetic changes induced by the transfected gene of yeast strains to the iron concentration variation the response.

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