Abstract: The calmodulin-domain protein kinases（CDPK）gene was cloned by RT-PCR from total RNA which was extracted from sporozoite of Eimeria tenella Gansu strain. Using the CDPK gene, the pGEX-CDPK recombinant plasmid was constructed. The recombinant plasmid was transformed into E.coli BL21 and highly expressed. The acquired fusion protein was purified by glutathione resin volume.The amount of fusion protein in total bacteria protein were 35.4%. The CDPK gene of Eimeria tenella Gansu strain was consisted of 1 470 nucleotides, encoding 490 amino acids. Compared with the CDPK gene of Eimeria tenella that reported in GenBank there are five mutation nucleotides and the homology of nucleotides is 99.6%, and there are five mutation amino acids and the homology of CDPK amino acids is 98%.