Cloning and Transformation of EeHKT1;4 Gene from Elytrigia elongata

The EeHKT1;4 gene was firstly cloned from Elytrigia elongata by RT-PCR technique with 1977 bp full-length cDNA encoding 1722 bp open reading frame (ORF) and 573 amino acids. The PCR fragment of EeHKT1;4 gene was inserted into the binary vector pBI121 and got the resulted expression vector, which named pBI121-35S-EeHKT1;4-Nos. The vector was further transformed into the agrobacterium EHA105, and then EeHKT1;4 gene was transferred into tobacco by the Agrobaterium- mediated genetic transformation method. The results showed that the target gene was inserted into the genomes of tobacco and expressed. Therefore, the transgenic tobacco (T0) plants overexpressing EeHKT1;4 gene were successfully obtained in this study. And EeHKT1;4 reduces Na+ concentration in the leaves of T0 plants, thereby plays a central role in protecting plant leaves from salinity stress. Keywords: Clone, EeHKT1;4 gene, Elytrigia elongata (Host) Nevski, plant expression vector, transformation, transgenic tobacco.