Construction of the wild-type and mutant interleukin13 expression plasmids in mammalian cells

Objective:To construct the expression plasmids of wild-type and mutant interleukin-13 expression plasmids in mammalian cells.Methods:The hGH-hIL13 fusion protein gene was amplified by fusion PCR.Both the PCR product and the vector pcDNA3.1(+) were digested by the Hind Ⅲ and EcoRⅠ.Then the PCR product was cloned in the eukaryotic expression vector of pcDNA3.1(+).The plasmids were constructed and transformed into E.coli competent cells DH5α.The positive clones were selected,and tested by sequencing.Results:The hGH-hIL13 fusion protein gene was in accordance with the design and correctly inserted into the vector proved by sequencing.Conclusion:The successful construction of the pcDNA3.1(+)/hGH-hIL13-a and pcDNA3.1(+)/hGH-hIL13-g plasmids lays the foundation for the wild-type and mutant interleukin-13 expression in mammalian cells.