The Tissue Specific Expression And Subcellular Localization Of AtJ70 In Arabidopsis

J-protein is a kind of protein which contains J-domain. Some J-proteins act as molecula chaperone. More than 90 J-proteins were identified in the genome of the model flowering plant Arabidopsis thaliana, but most J-protein’s function was unknown. AtJ70 is one of J-protein family members found in Arabidopsis thaliana. Its characteristic,localization and function have not been reported yet so far. In this project, tissue specific expression of AtJ70 gene was investigated using the GUS reporter gene and subcellular localization of this gene was studied using the GFP reporter gene.AtJ70 promoter was obtained using the method of PCR and the binary vector including the chimeric gene AtJ70 promoter: GUS was constructed. The binary vector was transformed into wild Arabidopsis and the transgenic lines were obtained .To determine the expreession of AtJ70 in different organs, the seedlings, flowers and siliques were GUS histochemical stained. The results indicated that except for stems, anthers and seeds, AtJ70 was expressed in roots、leaves、stigmas, filament and siliques. The expression of AtJ70 was the most strongest in vascular tissue and apical meristem. The results with leaves of different development stages showed that the expression of AtJ70 decreased obviously with senscence of leaves.Based on biologic informatics analysis, AtJ70 may be located in the nucleus. To confirm the location, we conducted genetic transformed experiment. Firstly, AtJ70 gene was cloned using the method of RT-PCR. Secondly, the binary vector including the chimeric gene AtJ70-GFP was constructed. Finally, the binary vector was transformed into wild Arabidopsis and the transgenic lines were obtained by screening with kanamycin. The root cells of the transgenic lines were viewed using Confocal. The result indicated that green fluorescence was distributed widely in whole cell in empty vector control line, but in the AtJ70-GFP transgenic line green fluorescence was found only in the nucleus. There is a big difference in contrast to the GFP control. The result indicated that AtJ70 was likely to be located in the nucleus. It was consistent with the analysis of the informatics.