RESUMEN
Polycystic ovary syndrome (PCOS) is the most common endocrine disorder affecting women of childbearing age, which features oligo- or anovulation, polycystic ovaries, hyperandrogenism and the related clinical signs, such as acne and hirsutism. At present, PCOS patients are considered to be in a long-time condition of chronic inflammation. It is reported that increased expression of inflammatory factors and/or increased levels of inflammation exist in peripheral blood, granulosa cells, follicular fluid, ovarian stroma, adipocytes and endometrial cells in patients with PCOS. Studies on the role of inflammatory factors in the pathogenesis of PCOS suggest that inflammatory factors may have an influence on the clinical outcome through affecting follicular development, androgen levels and so on.
RESUMEN
Objective: To express the fusion protein of hOct4 and cell penetrating peptides using prokaryotic expression systems, and to optimize its expression methods and observe the membrane penetrating ability of the fusion proteins. Methods: The pET-based prokaryotic expression system was constructed by genetic engineering, and the fusion plasmid was transferred into E. coli BL21(DE3) and Rosetta2(DE3). The protein was purified by Ni affinity chromatography and identified by Western blotting analysis. The penetrating ability of the Rhodamine-labelled fusion protein was investigated using BJ cells. Results: We successfully constructed pET21a(+)-hOct4-11R-His and pET21a(+)-EGFP-11R-His vectors. Fusion proteins hOct4-11-His and EGFP-11R-His were generated by transfering the plasmids into E. coli. The fusion protein was verified by Western blotting analysis and was detected in BJ cells. Conclusion: We have successfully generated EGFP-11R-His and hOct4-11R-His fusion proteins, and the proteins can effectively enter the BJ cells and locate around the nuclei.