Cloning And Functional-verify Of MnSOD Gene From Tamarix.androssowii

Tamarix.spp has high ability in resisting salt and drought enviroment .This research is to prove the function of MnSOD gene in Tamarix.spp by cloning MnSOD gene from Tamarix.spp,and constructing it in yeast expression vector pYES2,then transforming it by using budding yeast.The main results are:(1) The RACE technique was carried out in obtaining the cDNA sequence of MnSOD in T.androssowii,the full ORF has 699bp,which code 232 amino acids.The 5' end of the noncoding region has 40bp,and the 3'end of the noncoding region has 333bp.The bioinformatics methods were employed to analyse the protein.The protein's isoionic point is 7.10,and it's molecular weight is 26KD(2) The recombinant plasmid pYES-SOD was obtained by connecting MnSOD gene and yeast expression vector pYES2.Transform pYES-SOD into yeast fungus INVScl.Chose 4 INVScl (pYES-SOD) signal colonies randomly to prove MnSOD gene has been constructed in pYES2 vector through PCR.(3) In an attempt to investigate the expression of exogenous gene MnSOD in yeast by inducing INVScl (pYES-SOD) strain and INVScl (pYES2) strain. The result shows that INVScl (pYES-SOD) recombinant yeast can express exogenous gene by inducing. The highest level of the activity of SOD can be reached 2.064U/g-min.(4) The recombinant yeast INVScl (pYES-SOD) was treated with NaCL NaCO₃ NaHCO₃ drought rays.The result of this experiment reveals the stress-resistance of NVSc1(pYES-SOD) yeast was better than that of the control INVScl (pYES2) strain. That shows the SOD,expressed by MnSOD gene, protects the yeast cell when the yeast was treated with adversity stress. The result also proves the MnSOD gene,which came from Tamarix. spp ,has the function to improve the stress-resistance ability of eukaryote.(5) The methods of RT-PCR and Southern were used to detect the experiment. The result denotes that the treat with NaHCO solution and NaCl solution could increase the expression of MnSOD gene in Tamarix.spp. That shows the MnSOD gene in Tamarix.spp is related to salt resistance...