Cloning And Expression Of Gastric Cancer Related Gene S100P In E.coli As Well As Preparation Of Polyclonal Antibody

S100P, the differentially expressed gene was screened directly by fluorescent differential display reverse transcription polymerase chain reaction (DDTR-PCR) analysis from human gastric cancer and corresponding paratumor tissues in our laboratory. S100P is a member of the S100 family of genes and is located at 4pl6. Recent researches have showed that The S100P gene is significantly related to the development and advance of some malignant tumors. However the function of S100P is not very clear.We adopted the classical molecular biologic means, that is from gene to protein and then to antibody. It makes a foundation for clinically gaistric cancer detection for the highly pure and specific antibody. We first clone its full CDs length into the prokaryotic expression vector pET-22b(+),and the recombinant protein was expressed in the E.coli BL21(DE3)PlysS. Making use of the polyHis tag in the C-deminal of the recombinant protein ,we purified S100P protein. Polyclonal antibody against S100P was generated by immunizing mice with purified S100P recombinant fusion protein. Western blotting indicated that the antiserum against S100P could bind with the expressed recombinant protein specifically and as well as make a foundation of validate the relationship between S100P and gastric cancer.The results showed when diluted to 1:16000, the prepared polyclonal antibody had better immunoreactivity. It strongly testified the high purification of S100P protein as well as high specificity of polyclonal antibody. The study made a foundation of further studying S100P biological function in the development and advance of gastric cancer.