Cloning and Expression of Goat IFN-γGene and Preparation of Polyclonal Antibodies

To obtain the expression products of goat IFN-γ,total RNA was extracted from activated goat PBMCs,and cDNA was synthesized and used as template for PCR.The recombinant cloning vector was constructed.The sequence encoding mature peptide region was cloned by PCR and inserted into the expression vector pET-32 a,and the recombinant plasmid was transformed into the competent cell BL21(Codon Plus),followed by sequencing and analysis.After induced by IPTG,the expression products were and analysized by SDS-PAGE and purified by Ni-NTA.The purified protein was immunize rabbits to prepare polyclonal antibodies.Sequence analysis suggested that sequence encoding the mature peptide was 432 bp.SDS-PAGE indicated that the fusion protein was about 34.9ku,and a large amount of soluble protein could be expressed when induced with a total concentration of 0.3mmol/L/L IPTG for 6hat 30℃.The titter of polyclonal antibodies was 1∶106 detected by indirect ELISA.