Abstract: Objective To clone and express the C2 strain of Giardia lambia End-binding protein 1(gEB1) in E. coli. Methods Recombinant gEB1 protein was obtained, on which basis the primer was designed to clone gEB1 gene using C2 strain of Giardia lamblia as template and sequence reference to gEB1 gene by international standard WB strain The PCR product was cloned into prokaryotic expression vector pE-28α(+) with restriction enzymes Nco I and Xho I. The recombinant vector pET-28a(+)-gEB1 was transformed into E. coli host strain Rosetta(DE3), then the fusion protein was expressed by IPTG induction at 30℃ for 5 h. All products were tested and confirmed by SDS-PAGE and Western blot. Results The prokaryotic expression vector was successfully constructed, and the EB1 was highly expressed in E. coli Rosetta (DE3). SDS-PAGE and Western blot showed that the expressed product was about 29 kDa that was consistent with the defined value. Conclusion gEB1 gene has been successfully cloned and expressed in E. coli Rosetta(DE3), the recombinant protein can be used for further functional study and antibody preparation.

Key words: Giardia lambia, End-binding protein 1, Prokaryotic expression