| Arabidopsis thaliana, as a first completed genome sequencing of the plant, so far, there are still a lot of unknown functions to us to explore. With reverse genetics and the development of modern molecular biology techniques improved, these features unknown genes'locationing, the analysis of gene function, gene expression and regulation of research is increasingly gaining importance.This paper aims to study the unknown function on resistance and the location of Arabidopsis genome gene At4g32960. At4g32960 is in Arabidopsis chromosome 4, it,s full-length is 1563 bp, the gene encoding the protein sequence of about 264 amino acids (about 29 KD). Software forecasts indicate that it is not coding for the signal peptide, nor is it mitochondria or chloroplast protein. With the animal gene comparison, the gene may speculate as a transcription factor in plant dormancy process.First, we find the genome sequence from the web site called Tair, speculate there may be its promoter region by the analysis software Promoter2.0, then primers were designed, the promoter sequence was cloned and even linked to the appropriate vector pCAMBIA3301, after spending disseminated act transformed into Arabidopsis when it expresses in vivo, we have selected herbicide resistant seedlings because of the superiority of Bar gene of vector itself.Tthrough genetic testing on the report we can identify the histochemistry localization of the gene.We found that the gene location of the growth periods vary: during seedling period, expressed in the root position; during vegetative stage ,in the root guidance, Veiny, Trichome. And during reproductive growth, gene transfers to the root of the mature areas and anthers.Also, we constructed At4g32960-EGFP fusion gene expression vector, and disseminated by law into Arabidopsis flowers, we also have selected herbicide resistant seedlings and then through confocal microscopy detection of At4g32960 conducted an initial subcellular localization. After the fusion gene into yeast expression vector YEplac195 it entered into yeast cells and we used confocal microscopy for detection of At4g32960 subcellular localization, too. Two experimental results show that the gene expresses in the cytoplasm. Cloned At4g32960 and linked to pCAMBIA3301 to construct plant excessive expression vector, then transform it into Arabidopsis to obtain homozygous for the function study. And further the gene together with Histag was transformed into yeast expression vector YEplac195, excessive construction of yeast expression vector into the wild-type yeast cells K601. Screening positive clones by PCR, and we even carried out Western blot to further verified that the gene has expressed in vivo in yeast. Through the comparison of the growth of transgenic yeasts and yeasts of the wild-type under various stress conditions we speculate the gene,s function. By comparition we found that the growth in high-salt, high temperature, low temperature stress conditions, the growth of transgenic yeasts situation is obviously better than the wild-type yeasts, and in oxidative stress and low-salt conditions, the growth of the two kind yeasts do not have much difference, which indicates that the gene may be involved in the plant's adjustment of high-salt, temperature stress.At the same time we ordered At4g32960 gene mutant of Arabidopsis thaliana- salk-106519, and through detecting the expresstion of the gene we have got homozygous, then we plant homozygous mutant and wild-type Arabidopsis thaliana at the same time under the salt, H2O2, ABA, high temperature, low temperature stresses, etc. Training under the conditions, we found that the high temperature of the homozygous mutant,s growth is better than the wild-type while umder other conditions mutant and wild-type plants growth similar. Because of this we speculate that the gene may be involved in the stress regulation of Arabidopsis growth under high temperature, the results proved the yeast experiments of abrove.
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