Cloning the coding region of ODNaTx8 gene from Odonthobuthus doriae in E. coli

Background: Odontobuthus doriae is the most toxic Iranian yellow scorpion that is belong to the Buthidaefamily. Scorpion venome is a rich source of biologically active peptides which some of them are neurotoxins.ODNaTx8 gene coding a sodium channel blocker. Studies showed the roles of voltage-gated sodium channelblockers in neurological diseases such as apoplexy, cancer, MS and autism. In this work we cloned the codingregion of this gene in E. coli.Materials&methods: The cDNA encoding Odontobuthus doriae sodium channel blocker toxins(ODNaTx8) wasisolated from the library, replicated via PCR then this product and our vectors double digested with restrictionenzymes, then ligated and transferred to the E. coli bl21(DE3) through the expressional vector pET28.Results: We cloned ODNaTx8 gene into an expressional vector pET28 and transferred to E. coli as a host cell.Then we observed the recombinant clones and check them with clone-PCR and sequencing. For achievingrecombinant protein, induced the recombinant clones with IPTG, then detected our protein with SDS-PAGEtechnique.Conclutions: Recombinant protein of ODNaTx8 produced with cloning method. The importance of cloning thisgene is that it can produce the product of this gene and purify it from a large variety of proteins. By purifying andidentifying the product of the gene, it can be produced during the process of expression, producing an unlimitedamount of its product, which is far more advantageous and less costly than taking a large number of animalsamples, extracting poison from it, we can then isolate different proteins from it.

کلیدواژه ها:

Neurotoxins ، Cloning ، ODNaTx8 ، Sodium channel blockers ، Vectors