Cloning Of The Nitrate High-affinity Transporter Gene(TaNRT2;1) From Wheat And The Construction Of Fusion Protein

Genes of high 梐ffinity nitrate transporters have been cloned in higher plants recently. A full- length cDNA was isolated from Triticum aestivum by using RACE according to the known fragment of Thiticwn aestivum high affinity nitrate transporter . The cDNA ,denominated TaNRT2; I ,has a ORF of I 524bp and encodes a predicted protein of 507 amino acids residues (molecular weight 55.2kD ) with 12 putative transmembranes .A significant homology has been found between TaNRT2; 1 and the family of high-affinity nitrate transporters in other higher plants. We submitted this cDNA to Genbank and its accession number is AF3322 14. In order to acquire corresponding antibody to facilitate the following research of the cellular location of TaNRT2; I ,a fraction of TaNRT2; 1 , was constructed into the fusion protein expression plasmid , pGEX-2T and pET32a(+) ,respectively . The protein electrophoresis between recombinant plasmid and pure plasmid has distinct difference. Nucleic acid sequencing showed the translation fragment is right. But the fusion protein is still not obtained by now.