title: Cloning and expression of Neospora caninum dense-granule 7 in E. coli.
abstract: Neospora caninum is an obligate intracellular protozoan which induces abortion, still birth and neuromuscular disorders in cattle and is an important problem in dairy and beef industry worldwide. The dense granule protein 7 (GRA7) of N. caninum is an immune-dominant protein shared by both tachyzoite and bradyzoites. This study was conducted to produce recombinant GRA7 of N. caninum using a plasmid with high level of expression of this protein in E. coli. For this purpose, a segment of N. caninum DNA corresponding to GRA7 was amplified using PCR. After sequencing, this fragment was cloned into expression vector pMAL-c2X under the control of the lac promoter. Expression of this plasmid in E. coli strain TG1 was identified by western blotting. In this study, pMAL-c2X had a strong promoter to produce high level expression of NcGRA7. This result revealed that this recombinant protein with pMAL-c2X vector may be suitable for developing of diagnostic procedures.
authors.affiliation: Department of Parasitology, Veterinary Medicine Faculty, Shahid Chamran University, Ahwaz, Iran.
authors.forename: Marziye
authors.initials: M
authors.lastname: Kefayat
authors.affiliation: Department of Parasitology, Veterinary Medicine Faculty, Shahid Chamran University, Ahwaz, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Hamidinejat
authors.affiliation: Department of Microbiology, Veterinary Medicine Faculty, Shahid Chamran University, Ahwaz, Iran.
authors.forename: Masoud Reza
authors.initials: MR
authors.lastname: Seifiabadshapoori
authors.affiliation: Razi Vaccine and Serum Research Institute, Shiraz University, Shiraz, Iran.
authors.forename: Mohammad Mehdi
authors.initials: MM
authors.lastname: Namavari
authors.affiliation: Department of Parasitology, Veterinary Medicine Faculty, Tehran University, Tehran, Iran.
authors.forename: Parviz
authors.initials: P
authors.lastname: Shayan
authors.affiliation: Department of Clinical Sciences, Veterinary Medicine Faculty, Shahid Chamran University, Ahwaz, Iran.
authors.forename: Saad
authors.initials: S
authors.lastname: Gooraninejad
keywords: Cloning; E. coli; Expressing; NcGRA7; Neospora caninum