Prokaryotic Expression and Purification of Theileria annulata Recombinant Antigen Tams1-spag Genes

To study the purification method of the of fusion protein of Theileria annulata surface recombinant antigens and to obtain high purity products at low cost,different parameters were set in order to obtain the optimum of expression.The experimental parameters included OD600 nm,IPTG concentration induction time.Firstly,the target proteins were isolated by cutting the gel slices that contained the right bands which were stained by KCl solution.Secondly,the recycle protein were recovered by electrical elution and high centrifugation.Through SDS-PAGE and Western-blot,the experimental results indicated that the maximal quantity of expression of recombinant proteins was confirmed when IPTG concentration was 1mmol/Land OD600nm=1,the induced time was stay overnight.The two methods can obtain the same purified products and high purity.Western-blot assay indicated that the recombinant protein were not denaturated,and had good antigencity.