Cloning and Sequencing of Iranian Chicken Interleukine-2 Gene

Background: : IL-2 is a cytokine that plays an active role in the activation and mainte- nance of both acquired and innate immune defenses. It is also capable of improving the protective immune responses that are generated by conventional vaccines against avian pathogens in the poultry industry when used as an adjuvant. Objectives: The aim of this study was to extract and sequence Iranian chicken IL-2. Materials and Methods: In this study, genomic DNA was extracted from Iranian chick- ens. Total RNA was isolated by culturing harvested splenocytesand lysing them with Trizol reagent per the manufacturerʹs instructions. mRNA was isolated and converted to cDNA using reverse-transcriptase (RT) and specifc designed primers. Then, the PCR product was ligated into the PTZ57R/T plasmid (TA-cloning) and transformed into com- petent Top10 E. coli cells. Results: A unique 668-bp band was obtained after RT-PCR. Restriction enzyme digestion and colony PCR analysis and direct sequencing confrmed the existence of the desired gene in transformants. Conclusions: For frst time in Iran, the chIL-2 gene was successfully extracted, cloned, and transferred into E. coli. The efcacy of recombinant or DNA vaccines can be modu- lated by codelivery of this cytokine gene.