OVEREXPRESSION AND PURIFICATION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 ENV DERIVED EPITOPES IN ESCHERICHIA-COLI

In order to develop a reliable and inexpensive serodiagnostic method, a part of envelope gene of HIV-1, gp 120' and gp41' (HIV-1 enu a.a. 29 5-474 and a,a. 556-647) was cloned into a T7 expression vector (pET3d) , The fusion protein (gp120'-gp41') was overexpressed in Escherichia c oli, then purified to homogeneity by a simple gel filtration chromatog raphy, Western blot analysis and enzyme-linked immunosorbent assay (EL ISA) using the purified fusion protein showed a high sensitivity and a specificity for the detection of anti HIV-1 antibodies in testing hum an plasma, These results suggest that the expression scheme employing a direct expression vector and the rapid purification method are relia ble and applicable for obtaining a large quantity of HIV-1 env protein for diagnoses of HIV-1 infections.