Cloning and Expression of Chicken IFN-γ gene

The total RNA was abstracted from the concanavalin A(Con A) -activated spleen lymphocytes. A pair of specific primers was designed by Oligo software based on the sequence of Chicken interferon-gamma submitted by Digby in GenBank (accession No.U27465). A 500 bp DNA fragment was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated into T-easy vector for sequencing. The result shows that the fragment contained the complete open reading frame of CHIFN-γ gene. In comparison with GenBank data, the homology of the nucleotide sequence is 100% .The sequence was inserted into pGEX-4T-1 and the recombined expressing vector was constructed, then transformed into Escherichia coli BL21. which were further induced by IPTG and cultured,and a fusion protein was obtained with about 43 000 of molecular weight in SDS-PAGE. This result showed that the cloned CHIFN-γ gene was expressed in prokaryotic cells.