Cloning, Prokaryotic Expression and Protein Identification of eIF3h Gene from Ovis aries

Objective To clone gene of ovis aries eukaryotic translation initiation factor 3 subunit H and construct its prokaryotic expression vector,induce the expression and identify target protein in vitro.MethodFirstly,the ovis aries gene of eIF3h would be cloned from Ovis aries cDNA library using the primers based on eIF3h sequence.The prokaryotic expression vectors were transformed into expression host strain BL21,then after,IPTG was added to induce expression.SDS-PAGE and Western blotting assays were used to detect the expression of target protein.ResultIt successfully cloned eIF3h gene sequences of CDS.The length of the CDS was 1,059 bp.The Escherichia coli containing recombinant vector expressed inclusion body proteins of 40kD after induction by IPTG at 37°C.ConclusionThe construction of recombinant plasmid is successful,which has provided the basis for further research of eIF3h molecule.