Cloning, Expression and Purification CEL I Endonuclease Enzyme from Celery Plants

CEL I، is isolated from celery and the first eukaryotic nuclease is known to have cleavage DNA with high specificity at sites of the two DNA strands in a heteroduplex DNA molecule. The aim of this study was cloning، expression and purification CEL I endonuclease and a simple method for the detection of mutation s. CEL I mRNA was used for RT-PCR. CEL I cDNA was synthesized and cloned into T-vector pTZ57R، then CEL I subcloned into pET32a expression vector. Recombinant plasmid was transformed into BL2I bacterial cells. Expression of recombinant plasmid was analyzed by western blot technique. Protein was purified by affinity chromatography. In first study، Recombinant enzyme was active in mutation detection in Product PCR normal and mutant BRCA1 gene. he recombinant (pET32a /CEL I) was successfully expressed in BL2I cells. Western blot analysis showed that the successfully expressed CEL I in the cells transfected. The first study showed that recombinant enzyme is kno wn to nick mismatch sites of the two DNA strands in a heteroduplex DNA molecule. The expression of CEL I in BL2I cells was strong. In vitro، purification protein was successful. Recombinant nucleases CEL I known to nick mismatch sites in a heteroduplex DNA molecule.