Proteome And Transcriptome Maps Analysis Of Transformed Yeast With MxIRT1 Or OsNramp1

MxIRT1 and OsNramp1 were metal transporters cloned from M. xiaojinensis Cheng et Jiang and rice roots respectively under iron-deficient conditions. Both of MxIRT1 and OsNramp1 can complement iron uptake activity of Saccharoomyces cerevisiae mutant strain DEY1453 (fet3fet4). This work is aimed to research the pathway induced by the two genes in the level of proteome and transcriptome maps.Part one: Proteome analysis of transformed yeast with IRT cultured under iron-sufficient and iron-deficient conditions, transformed yeast with Nramp and empty-vector cultured under normal condition, transformed yeast with IRT Nramp and empty-vector under the normal or iron-deficient conditions. Using 2-DE technique more than 1500 spots were isolated per gel. After the in-gel digest and MALDI-TOF-MS, 8, 9 and 11 proteins with difference were identified respectively under the Mascot research. Most of the proteins were associated with the cytoskeleton, chaperone, metabolism, protein biosynthsis, cell resistance, vesicle transport and signal conduction et al.Part two: Transcriptome maps analysis of transformed yeast with IRT Nramp and empty-vector under the normal iron conditions. Extract the yeast total RNA and labeled biotin in the way of reverse-transcription, and then analyze gene expression at transcriptional levels by Microarray and software in the 10928 individual gene sequences. 1200 genes were up-regulated expression while 1140 down-regulated in the strain of transformed yeast with IRT. 290 genes were up-regulated expression while 108 down-regulated in the strain of transformed yeast with Nramp. We analyze the prominent 27 up-regulated genes and 45 down-regulated genes, most of the up-regulated genes were related to zinc-finger, RNA binding, regulator or inhibitor and induced by high-osmotic while the down-regulated genes were related to vesicle transport, cell communication, cell cycle and biosynthesis of proteins.We hypothesize that in the transformed yeast, cells can form diversiform biology pathway in response to the iron concentration to maintain normal growth.