Subcellular Localization And Functional Analysis Of OsNramp1 Gene

Plants couldn't get enough and available iron for its low solubility in soil,which lead to a universal and widespread human health problem-iron deficiency.Many plant basic functions need plant transporter to carry out ranging from metal absorption to metal sequestration and storage.Nramp,as a super metal transporter and membrane integral protein family,plays a very important role in metal up-take,transportation and homeostasis.The results of Real-time PCR suggest that the expression of OsNrampl is up-regulated heavily by 14 times under iron deficiency,which illuminates that this gene may be involved in the uptaking and transportation of iron.Positive and homozygous sense,antisense and only-vector transformed tobacco were obtained by leaf disc method.Then I have done my research in these ways,as follows:1.Through the phonotype observation and photosynthesis indexes testes with the transformed tobacco we found OsNRAMP1 have not obvious effects to plant growing.We tested the iron content of the transformed tobacco parts and seeds,and found OsNRAMP1 can promote some metal ions' accumulation in vegetative parts and seeds.2.With the DIET technology we tested the relationships of OsNRAMP1 and Ca²⁺,Cd²⁺,H⁺, but no remarkable signal was detected.3.The biology bioinformatics software predicted OsNRAMP1 is a plasma membrane-located membrane integrated protein.And we found OsNRAMP1 is mainly localized on ER and cell membrane with the help of FM-4-64 and Blue-White.The same time we observed some special vesicular in cytoplasm and spots with GFP signal gathering,which may come from the ER and transport to cell membrane playing its function.4.We detected obvious hybridization signal with the extracts of transformed E.coli which over-expressing OsNRAMP1.And taking the protein purification with the His marker provides a base for lately antibody preparation.