|
|
Avicenna Journal of Medical Biotechnology، جلد ۳، شماره ۲، صفحات ۶۷-۷۸
|
|
|
| عنوان فارسی |
|
|
| چکیده فارسی مقاله |
|
|
| کلیدواژه‌های فارسی مقاله |
|
|
| عنوان انگلیسی |
Expression and Single-step Purification of GRA8 Antigen of Toxoplasma gondii in Escherichia coli |
|
| چکیده انگلیسی مقاله |
Diagnosis of Toxoplasma gondii (T.gondii) infection is of great medical importance especially for pregnant women and immunosuppressed patients. Numerous studies have shown that the recombinant production of several tToxoplasma antigens, including dense granule antigens (GRAs) has a great potential as diagnostic reagents. Previous studies reported expression of amino terminal GRA8 protein in fusion with large tags. In the present study, we produced truncated GRA8 (TGRA8GRA8), excluded from the signal peptide and C-terminal transmembrane domain, with a short fusion tag in Escherichia coli. (E.coli). TGRA8GRA8 was purified using an optimized single-step Iimmobilized Mmetal ion Aaffinity Cchromatography (IMAC). The purity and yield of TGRA8GRA8 was highest at pH= 9.25. At this pH, 13.6 mg of TGRA8GRA8 was obtained with the purity of 97.97%. Immunogenicity of the protein was evaluated in Western blot analysis showing the serum sample from a rabbit immunized with TGRA8GRA8 recognized a single antigen of T. gondii tachyzoite at the expected molecular weight of native GRA8. To diagnosis acute tToxoplasma infection in pregnant women, an indirect immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) was developed using TGRA8GRA8 resulting in a test specificity and sensitivity of 97.1% and 60.6%, respectively. These results demonstrated that immunogenic TGRA8GRA8 can be produced in fusion with a short tag and purified near to homogeneity using an optimized IMAC. TGRA8GRA8-IgM-ELISA was useful for detection of acute tToxoplasma infection. |
|
| کلیدواژه‌های انگلیسی مقاله |
|
|
| نویسندگان مقاله |
|
|
| نشانی اینترنتی |
http://www.ajmb.org/En/Article.aspx?id=58 |
| فایل مقاله |
اشکال در دسترسی به فایل - ./files/site1/rds_journals/133/article-133-376201.pdf |
| کد مقاله (doi) |
|
| زبان مقاله منتشر شده |
en |
| موضوعات مقاله منتشر شده |
|
| نوع مقاله منتشر شده |
|
|
|
|
برگشت به:
صفحه اول پایگاه |
نسخه مرتبط |
نشریه مرتبط |
فهرست نشریات
|