Identitfication And Preliminary Functional Analysis Of OsAPX1 Gene In Dongxiang Wild Rice

Rice is not only the most important crop in the world, especially for China, but also a promising model plant for plant molecular and functional genomics research. Dongxiang wild rice( Oryza rufipogon Griff), as the most northern and cold tolerant common wild rice in the world as so far, has a strong ability to resist low temperature in seedling stage and heading stage. Therefore, identifying new cold stress-related genes and understanding their functions will have an important practical significance in improving the low temperature tolerance to adverse growing conditions and variety upgrading.In this study, a novel function-unknown gene was discovered through expression profiling and bioimformatics analysis. A gene Os APX1 with an ORF of 750 bp encoding Os APX1 protein was screened and identified from c DNA library of the Dongxiang Wild rice. Bioinformatic analysis of the protein showed that this protein is mainly composed of polar amino acids exhibiting 99% homology with that of common rice.1. Successfully screened from Dongxiang wild rice root c DNA library encoding Os APX1, ORF of 750 bp, with ordinary rice up to 99% homology.2. Target gene Os APX1 and p ET28 a vector was ligated and transformed into DH5α, the recombinant vector construct p ET28-APX1, analysis showed that the target gene in the correct ORF inserted into the vector.3. Clones were screened positive DH5α recombinant vector is transformed into th e expression BL21. Screened positive recombinant bacteria, induced with IPTG, the overexpression of proteins by SDS-PAGE electrophoresis analysis showed that Os APX1 was expressed successful in BL21, the protein molecular weight was about 27 k Da.4. After optimization of the experimental conditions, to give a higher content of soluble protein. The protein in the non-deformed condition by Ni-NTA Superflow column obtained a single protein band.5. Of wild rice plants Os APX1 gene expression levels at different times and in different tissues, found Os APX1 heading and maturity stages of gene expression in the amount of enhancement; in floral organs and maturity of the strongest expression of fruit.6. Under different abiotic stresses, Os APX1 expression was raised, wich was a stress-related gene; Os APX1 played a vital role in the anti-stress gene networks.