Cloning and expression of the gene fragment encoding chicken MHCIIβ chain and its antibody preparation
The gene fragment of chicken MHC class II beta chain from spleen lymphocytes was cloned by RT-PCR using a pair of primers.It was 798 bp in length and showed 95% similarity to the previously identified chicken MHCⅡ beta at nucleotide level.The full length chicken MHCⅡ beta chain cDNA was cloned into pGEX-4T-1.The recombinant plasmid,namely PGEX-4T-chMHCⅡβ,was transformed into Escherichia coli BL21 and GST-β fusion protein was induced to express.A MW of the fusion protein was about 53 000 as analyzed by SDS-PAGE.The purified fusion protein was used to immunized mice and the specific antibody was observed,which was identified further in immunodiffusion and ELISA.The results indicated that this expressed fusion protein had favorable antigenicity.