Studies on expression and purification of FHIT/TAT fusion protein and preparation of its polyclonal antibody
AIM: To prepare fusion protein of fragile histidine triad (FHIT)gene with HIV-TAT protein transduction domain and to acquire rabbit polyclonal antibody against the fusion protein so as to lay a good foundation for further research on the function and mechanism of FHIT gene in hepatocellular carcinoma. METHODS: Prokaryotic expression vector of FHIT gene was constructed with HIV-TAT protein transduction domain and its optimal expression conditions were investigated. FHIT/TAT fusion protein was acquired by E. coli expression system and subsequent purification process and rabbits were immuned with the purified fusion protein to acquire polyclonal antibody against it. RESULTS: The expression vector of FHIT/TAT fusion protein was successfully constructed and the optimal expression conditions were determined. The target protein's accurate expression was confirmed by Western Blot analysis. The purity of FHIT/TAT fusion protein was nearly 70% and higher titer rabbit polyclonal antibody against FHIT/TAT fusion protein was acquired. CONCLUSION: Prokaryotic expression system of FHIT/TAT fusion protein with high transduction efficiency has been successfully established and higher liter rabbit polyclonal antibody againat FHIT/TAT fusion protein has been acquired,which lays a good foundation for the function and mechanism research of FHIT gene on hepatocellular carcinoma,especially on the gene therapy and prevention of hepatocellular carcinoma.
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万方数据
维普
