Cloning of human interleukin-2 gene in mammalian expression vector

The human interleukin-2 gene was amplified by reverse transcriptase polymerase chain reaction with M-MuLV reverse transcriptase enzyme from the total RNA of human peripheral blood lymphocytes. Taq DNA polymerase enzyme was used to amplify the gene as it adds ‘A’ overhang in the PCR product. This ‘A’ tailed product was cloned in TA cloning vector pTargeT and transformed in E. coli DH5á cells. The recombinant plasmid containing the human interleukin-2 gene insert in right orientation was selected after characterization using restriction enzyme analysis, directional PCR and nucleotide sequencing.