中国生物工程杂志

Aim The recombinant Human retinal pigment epithelium-derived factor（PEDF）protein to be obtained and the angiogenesis of the rPEDF to be identified. Method PEDF gene was amplified by PCR and cloned into pET32a, rPEDF protein was expressed in E coli BL21 and confirmed by SDS-PAGE and Western-blot. The rPEDF was purified by Ni-NTA on denature condition. The concentration of the rPEDF was determined by Bradford method. The angiogenesis of the rPEDF was determined by chick chorioallantoic membrane (CAM) method. Results The expression plasmid pET32a-PEDF was constructed successfully .The rPEDF was expressed stably efficiency in E coli BL21. The results of the CAM experiment showed that the rPEDF had notable angiogenesis effect in the concentration 0.4、0.04 ng/ml, but had no effect in 4 ng/ml. Conclusion: The PEDF gene was cloned and expressed efficiency, the angiogenesis of the rPEDF to be identified and the activity was worked in certain range. The results can facilitate studying its function and spreading its application.