TY - JOUR ID - 6888 TI - Expression of Recombinant 3-Beta Hydroxysteroid Dehydrogenase Protein in E. coli JO - Iranian Journal of Biotechnology JA - IJB LA - en SN - 1728-3043 AU - Vallian, Sadeq AU - Farzaneh, Farzin AD - Division of Genetics, Department of Biology, Faculty of Science, Isfahan University. Isfahan, I.R. Iran AD - Molecular Medicine Department, King´s College School of Medicine and Dentistry, King´s College, London, UK. Y1 - 2003 PY - 2003 VL - 1 IS - 2 SP - 109 EP - 114 KW - 3-beta hydroxysteroid dehydrogenase KW - Recombinant protein KW - autoantibody DO - N2 - 3-beta hydroxysteroid dehydrogenase (3BHSD) is secreted by the cortex of adrenal gland functioning instress conditions. The gene encoding the 3-beta hydroxysteroid dehydrogenase protein was PCRamplified from a λgt11 cDNA library using specific primers. The amplified PCR product was then cloned into pGEX-4T-1 expression vector under Ptac promoter and the expression of the enzyme was examined in E. coli (BL21). Upon optimization of the expression condition, the enzyme was produced as a glutathione S-transferase (GST) fusion protein, which was purified by affinity chromatography using glutathione sepharose column. The GST part was then removed by selective proteolytic digestion with thrombin. The purified recombinant enzyme could be used in construction of diagnostic kits for screening the patients with premature ovarian failure (POF) for the presence of autoantibodies against 3BHSD as an important molecular target. UR - https://www.ijbiotech.com/article_6888.html L1 - https://www.ijbiotech.com/article_6888_9d31f51e8df9975184973d56da81cbd9.pdf ER -