Cloning, expression, purification and identification of recombinant human C22orf37 fusion protein
AIM:To clone,express and purify C22orf 37 fusion protein in E. coli. METHODS: A 510 bp of human C22orf37 gene fragment was obtained by PCR method from human leukyocyte cDNA library and cloned into pQE30 vector,a His fusion expression vector. The recombinant plasmid was transformed into E. coli DH5α and induced to express fusion protein His-C22orf37 with IPTG. The products expressed in E. coli were analyzed by SDS-PAGE and identified by Western blot. The interesting protein was purified by Ni-NTA affinity chromatography and the purity of the purified protein was analyzed using the Primer Premier software. RESULTS:The recombinant plasmid pQE30-C22orf37 was constructed and the fusion protein was expressed in DH5a at a high level. SDS-PAGE showed that its molecular mass was about 18 kD. Recombinant protein was purified by Ni-NTA purification column and the purity of the fusion protein was about 80%. The His-C22orf37 fusion protein showed a good binding ability to anti-His antibody specifically. CONCLUSION: Our successful expression and purification of His-C22orf37 protein lay a basis for the production of anti-C22orf37 monoclonal antibody and further studies of the tissue distribution and function of C22orf37.
human C22orf37 geneprokaryotic expressionrecombinant fusion proteins/isolation & purification
NETL
NSTL
万方数据
维普
