| Abstract:Essential protein with sulfite reductase activity required in assimilatory sulfate reduction pathway during both primary and secondary metabolism. In this thesis, we have studied sulfite reductase and its sequence,and have got some results as follows:1. Cloning of the ZmSR and sequence analysis. We have got the full length of ZmSR from inbred lines of maize.chang7-2.zheng58.qi319and so on by Nested PCR. we have found that ZmSR belongs to the super families multi-domains. l-54aa is a transit peptide.import signal into chloroplast stroma, but exact processing site is unknown; And51-635aa is ferredoxin-sulfite reductase mature protein. But, exact N-terminus is unknown. Besides, there are two NIR_SIR_ferrs Region.The Amino acids sequence of maize has a high homology with the Amino acids sequences of other crops.2.The biochemical characteristics of ZmSR was studied.After constructing the recombinant plasmid pET30-SR and pEASY-E1-SR and transferring the pasmid into the competent BL21(DE3) cells, approximately a70kDa fusion protion was induced by addition of Isopropyl-beta-D-thiogalactopyranoside(IPTG)during cultivation in pEASY-E1-SR plasmid.3.The expression pattern of ZmSR under different stresses.The maize of zheng58was analysed by real time PCR after treatement with heat. cold. PEG and so on.4.Response mechanism of ZmSR which was treatment by SO2was studied with transgenic technology. The atsr by T-DNA insertion in Arabidopsis thaliana were treatment with SO2, The results showed that the atsr are sensitive to SO2and The degree of injury in atsr is higher Wild-type (WT) Therefore, the recombiant plant expression vectors with35S strong promoter was constructed and were transferred into Agrobacterium GV3101. Transgenic Arabidopsis thaliana with the method of Floral-dip.The seeds harvestded were analyzed by PCR.(WT:ZmSR is two Lines, atsr:ZmSR is ten Lines)... |
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