The Preliminary Study Of The Yeast Gene FUR4 Ardabidopsis Transgenic Line And At5g03555

The FUR4 protein is a uracil permease that mediates the uptake of uracil, but does not transport other natural pyrimidines such as cytosine, thymine, or uridine in S. cerevisiae. It is localized to lipid microdomains of the plasma membrane. FUR4 functions as a symporter by actively translocating uracil with the co-transport of protons, and the prodrug 5-fluorouracil (5-FU) enters the cells via FUR4, too. fur4 mutant is not sensitive to 5-FU. Overexpressing FUR4 is toxic in cells.To futher study the function of FUR4 protein, the FUR4 protein and its homologous gene At5gO3555 were mainly studied in Arabidopsis thaliana in this paper. The results confirmed that the FUR4 gene was successfully transformed in to Arabidopsis and stable genetic. The subcellular localization of FUR4 protein located in cell membrane and nucleus. Further analysis the phenotypes of FUR4 transformant, the results showed that anthers dehiscence were abnormal and pollens were less activity than wild type. And their siliques lengths were shorter, too. That led to reducing of seed rate or even completely abortion.The At5g03555 which shares 26% homology in protein sequence of FUR4 was researched. Reverse transcription-PCR analysis andβ-glucuronidase staining showed that At5g03555 expressed in different organs. However, the expression was much stronger in flowers and siliques. The subcellular localization of At5g03555 protein was found to be localized in the cell membrane and nucleus, which was the same as FUR4 protein. A T-DNA insertion mutant of At5g03555 was also analyzed. The phenotypes of At5g03555 mutants were undistinguishable from those of wild-type plants under normal growth conditions. And a comlementary by FUR4 gene was abtained. Under different stresses, mutant seeds germinated faster than wild type, while the complementary was similar to wild type. These data indicate that the features of FUR4 and At5g03555 were similar which may be relater to osmotic and ionic stress. They may also involve in the regulation of seed germination.It can study the mechanism of transmembrane transporters by FUR4 protein and At5g03555 protein in higher plant, which is helpful for the genetic disease which are coursed by lose function of transport system and maybe a perfect new target of therapy and agriculture intermediary.