Expression and functional study of green fluorescent protein-hemagglutinin A-cervical cancer fusion protein in vitro / 中国药学杂志

ABSTRACT

OBJECTIVE: To prepare green fluorescent protein-hemagglutinin A-cervical cancer (GFP-HA-PTP) fusion protein trageting HPV transformed cervical cancer cells with endosome escape capability, and further investigate its penetrating ability for cervical cancer cell lines. METHODS: pET15b-GFP-HA-PTP expression plasmid was constructed, and E. coli BL21 (DE3) was transformed with the plasmid, then the yielded GFP-HA-PTP fusion protein was purified by Ni-NTA His-Bind resin and conformed by Western blotting assay. Specific penetrating analysis of GFP-HA-PTP was performed under fluorescence microscopy. RESULTS: GFP-HA-PTP fusion protein was highly expressed in BL21 cells. Western blotting analysis result showed that the fusion protein was expressed correctly. The fusion protein can selectively penetrate into cervical cancer cell lines Siha and Caski with endosome escape efficiently. CONCLUSION: GFP-HA-PTP can specificly penetrate into cervical cancer cell lines after endosome escaping with high efficiency.