Construction and expression of recombinant ovarian cancer gene ScFv in pichia pastoris

Objective To obtain ScFv gene from ovarian cancer patients with gene recombinant technique and express it into pichia pastoris in order to provide experimental basis of application of ScFv in diagnosis and treatment of ovarian cancer.Methods The total RNA of lymphocytes cell was extracted from ovarian cancer patients by TRIZOL.VH and VL gene were obtained by SOEingPCR.pPICZa/ScFv was contructed and transformed into X-33 cells.The expression was induced by pichia pastoris.The engineer bacteria with high effective expression of ScFv was screened.Results ScFv gene was constructed successfully.ScFv gene was about 700 bp and protein was induced by pichia pastoris and it was identified that its molecular weight was about 26 000.Conclusion ScFv gene,pPICZα/ScFv and pichia pastoris engineer bacteria with high effective expression of ScFv are obtained.