Gene Cloning And Transformation Of Antioxiditant Enzymes In The Chloroplast Of Wheat

In order to obtain double transgenic tobacco plants with over-expressing SOD and APX .We have done some jobs about the cloning and transformalion of SOD and APX from wheat (vanct> Hanfcng 9703). The main results were as follows:1. We designed a pair of primers according to pulishcd Wheat Cu/Zn-SOD sequence and obtained a cDNA fragment about 8()()bp.Through analysis and nuclcotidc BLAST of this sequence, we know that it is the target sequence which shares 97.6% identity with published sequence in Gcncbank. But to our surprise, it is 80bp shorter than published sequence in the 3' end .2. Sense and anti-sense SOD expression vectors under control of 35Spromoter were constructed and transformed them into tobacco(Nicotiana tabacnm L.). Through PCR and Southern detection . it is suggested that we have obtained sense and anti-sense transgenic plants.3.According to conserved amino acid sequences of APX. we designed a pair of primers and obtained a cDNA fragment about 680 bp from Wheat leaves through RT-PCR. It shares high identity with thylakoid-bound APX genes. We registered it in Genebank . (F387739) .4.We aiso cloned 5' and 3" end sequences of the gene. Further a full-length cDNA sequence was obtained through 5' and 3 RACE-PCR.