CVN (Cyanovirin-N) is a highly potent anti-HIV protein. Cyanovirin-N homology (CVNH) family is a homologous protein to CVN

which exhibits a common fold structure similar to CVN and possess a highly conserved anti-HIV CVN domain. In this study

DNA encoding sequences of the wildtype and mutant CtCVNH from Ceratopteris thalictroides were separately cloned into eukaryotic expression vector pPICZαA. All wCtCVNH and mutated mCtCVNH genes produced two bands corresponding to 22 000 and 24 000

which were validated by western blot. The proteins were purified by ultra-filtered centrifugation and Ni-NTA affinity chromatograph. Glycosylation assay further confirmed that no N-glycosylation site occurred in wCtCVNH protein. The possible reasons were ascribed to incomplete cleavage of the α-factor signal peptide sequence in vector pPICZαA or some other post-translational modifications rather than N-glycosylation. The study laid a solid foundation for further exploring the pharmacokinetics and pharmacodynamics of CtCVNH in future.