title: Expression of recombinant human coagulation factor VII by the Lizard Leishmania expression system.
abstract: The variety of recombinant protein expression systems have been developed as a resource of FVII gene expression. In the current study, the authors used a novel protein expression system based on the Iranian Lizard Leishmania, a trypanosomatid protozoan as a host for expression of FVII. Plasmid containing cDNA encoding full-length human FVII was introduced into Lizard Leishmania and positive transfectants were analyzed by SDS-PAGE and Western blot analysis. Furthermore, biological activity of purified protein was detected by PT assay. The recombinant strain harboring a construct was analyzed for expression of FVII at the mRNA and protein level. Purified rFVII was obtained and in order to confirm the purified compound was in fact rFVII. Western blot analysis was carried out. Clotting time in PT assay was reduced about 30 seconds with the purified rFVII. In Conclusion, this study has demonstrated, for the first time, that Leishmania cells can be used as an expression system for producing recombinant FVII.
authors.affiliation: Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Sina
authors.initials: S
authors.lastname: Mirzaahmadi
authors.forename: Golnaz
authors.initials: G
authors.lastname: Asaadi-Tehrani
authors.forename: Mojgan
authors.initials: M
authors.lastname: Bandehpour
authors.forename: Nooshin
authors.initials: N
authors.lastname: Davoudi
authors.forename: Leila
authors.initials: L
authors.lastname: Tahmasbi
authors.forename: Nahid
authors.initials: N
authors.lastname: Hosseinzadeh
authors.forename: Hasan
authors.initials: H
authors.lastname: Mirzahoseini
authors.forename: Kazem
authors.initials: K
authors.lastname: Parivar
authors.forename: Bahram
authors.initials: B
authors.lastname: Kazemi
mesh_terms: D000818:Animals; D001780:Blood Coagulation Tests; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D005167:Factor VII / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D056945:Hep G2 Cells; D006801:Humans; D007891:Leishmania / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D008116:Lizards / Q000469:parasitology; D008958:Models, Molecular; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D005167:Factor VII