Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Two-dimensional gel electrophoresis and immunoblot analysis of Neospora caninum tachyzoites | J. Vet. Sci. 2004 5 2 139-145 JOURNAL OF Veterinary Science Two-dimensional gel electrophoresis and immunoblot analysis of Neospora caninum tachyzoites Eung-Goo Lee Jae-Hoon Kim1 Yong-Seung Shin Gee-Wook Shin Yong-Hwan Kim Gon-Sup Kim Dae-Yong Kim2 Tae-Sung Jung Myung-Deuk Suh Institute of Animal Medicine College of Veterinary Medicine Gyeongsang National University Jinju 600-701 Korea Department of Veterinary Medicine Cheju National University Jeju 690-756 Korea Department of Pathology College of Veterinary Medicine Seoul National University Seoul 151-742 Korea Identification of expressed protein profiles and antigenic determination are some of the most challenging aspects of proteomics. Two-dimensional gel electrophoresis 2-DE combined with immunoblot analysis were employed to study the N. caninum proteome. Protein sample preparation was carried out by first conducting sonication followed by adding lysis buffer containing 7M urea plus 2M thiourea to the purified tachyzoites in order to complete disruption. A total of 335 differentially expressed protein spots were detected using pH 4-7 IPG strip 7 cm that were run in a 56 kVh isoelectric focusing IEF system. Of the spots analyzed 64 were identified as antigenic spots on immunoblot profile. Major antigenic spots appeared at 65 kDa pl 51 kDa pl 38 kDa pl 33 kDa pl 29 kDa pl and kDa pl were observed to be significantly distinct compared to the rest of the antigenic spots. The results indicate that combination of 2-DE and immunoblotting methods were thought as very useful tools in defining both proteins and antigens of N. caninum tachyzoites. Additionally present 2-DE profiles may be valuable in further proteomic approaches and study of the pathogen. Kewords Neospora caninum two-dimensional gel electrophoresis 2-DE immunoblot Introduction Neospora caninum N. caninum is an obligate cystforming intracellular protozoan Apicomplexa parasite of animals 10 11 . It was usually misdiagnosed .
