Abstract: Objective To clone, express and purify recombinant protein C7orf69 in vitro, and to analyze its bioinformatics by prediction software. Methods Gene C7orf69 was amplified by polymerase chain reaction (PCR) and inserted into prokaryotic expression plasmid. Then the plasmid was transformed into E. coli BL21, induced by IPTG, identified by SDS-PAGE and Western blot, and purified under native or denaturing conditions by Ni-NTA resin. Results The right sequenced gene C7orf69 cotaining a natural variation was obtained, and the recombinant protein was highly expressed in the form of inclusion body. Purified recombinat protein was obtained under native conditions by Ni-NTA resin. The bioinformatics analysis by prediction software indicates that there are PKC and CK2 phosphorylation sites and a myristylation site in the protein C7orf69 including the signal peptide. Conclusions C7orf69 was successfully cloned, expressed and purified. These jobs lay solid foundation for the further study of its functions.