National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304310420121001Microbial cell surface display; its medical and environmental applications2312397171ENVida TafakoriDepartment of Molecular Genetics, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O.Box 14965/161, Tehran, I.R. Iran.Ibrahim TorktazDepartment of Biotechnology, Faculty of Advanced Science and Technologies,
University of Isfahan, 81746-73441, Isfahan, I.R. Iran.Mohsen DoostmohammadiDepartment of Biotechnology, Faculty of Advanced Science and Technologies,
University of Isfahan, 81746-73441, Isfahan, I.R. Iran.Gholamreza AhmadianDepartment of Molecular Genetics, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O.Box 14965/161, Tehran, I.R. Iran.Journal Article20121001Cell-surface display is the expression of peptides and proteins on the surface of living cells by fusing them to<br />functional components of cells which are exposed to the environment of cells. This strategy can be carried<br />out using different surface proteins of cells as anchoring motifs and different proteins from different sources<br />as a passenger protein. It is a promising strategy for developing novel whole cell factories. Surface engineered<br />cells have many potential uses ranging from medical to environmental applications. This review focuses on different strategy and applications of microbial surface display. <br />Displayhttps://www.ijbiotech.com/article_7171_54a154fd2b34e89985489edeaf46bb3a.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304310420121001Production and evaluation of polyclonal rabbit antihuman p53 antibody using bacterially expressed glutathione S-transferase-p53 fusion protein2402487180ENMozhgan RastiRecombinant Protein Laboratory, Department of Biochemistry, Medical School, Shiraz University of Medical
Sciences, P.O. Box 7134-845794, Shiraz, I.R. Iran.Zahed KhatooniRecombinant Protein Laboratory, Department of Biochemistry, Medical School, Shiraz University of Medical
Sciences, P.O. Box 7134-845794, Shiraz, I.R. Iran.Zoherh Mostafavi-PourRecombinant Protein Laboratory, Department of Biochemistry, Medical School, Shiraz University of Medical
Sciences, P.O. Box 7134-845794, Shiraz, I.R. Iran.Journal Article20121001p53 is a key tumor suppressor gene that is targeted for inactivation during human tumorigenesis. In this study, we produced and characterized polyclonal antihuman p53 antibody. The cDNA encoding the complete<br />human p53 protein was cloned into pGEX-4T-1 and expressed in Escherichia coli as a fusion protein with Schistosoma japonicum glutathione S-transferase (GST). The rabbits were immunized with the purified<br />p53 recombinant protein. The obtained antisera were purified to increase the specificity of recognition. The<br />sensitivity and specificity of the produced antibody was analyzed by enzyme-linked immunosorbent,<br />immunoblot, immunofluorescence and chromatin immunoprecipitation assays. Enzyme-linked immunosorbent assay showed that immunization with purified GST-p53 produced the high titer (1:10000) polyclonal antibodies with high specificity. Anti-p53 antibody allowed the sensitive detection of native p53 protein in immunoblotting, immunofluorescence and chromatin immunoprecipitation assays. Our results showed that anti-GST-p53 antibody provides a good means for studying the p53 expression pattern and its binding ability to other proteins in tumors.<br /><br />https://www.ijbiotech.com/article_7180_c389df12c31ca6d18c3d23bb57b226ed.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304310420121001Cloning, expression, purification and immunoreactivity analysis of gag derived protein p17 from HIV-1 CRF35 in fusion with thioredoxin from human subjects2492547177ENMohammad AskariDepartment of Medical Biotechnology, Faculty of Allied Medical Sciences, Tehran University of Medical Sciences, P.O. Box 1449614535, Tehran, I.R. Iran.Fazel GorjipourHIV Molecular Research Laboratory, Division f Virology, Department of Pathobiology, School of Public Health and Health Research Institute, Tehran University of Medical Sciences, P.O. Box 19395-5731, Tehran, I.R. Iran.Zohreh SharifiBlood Transfusion Research Center, Institute for Research and Education in Transfusion Medicine, P.O. Box 14665-1157, Tehran, I.R. Iran.Mohammad Morad FarajollahiDepartment of Medical Biotechnology, Faculty of Allied Medical Sciences, Tehran University of Medical Sciences, P.O. Box 1449614535, Tehran, I.R. Iran.Journal Article20121001So far, recombinant antigens of HIV-1, the etiologic cause of Acquired Immunodeficiency Syndrome (AIDS), have been widely used for the diagnosis and vaccine development. P17 or the matrix protein formed by the proteolytic cleavage of gag is strongly antigenic and is as conserved and immunogenic as p24. In some cases, antibodies to p17 are more prevalent than antibodies to p24 and the decline in the level of p17 antibodies is an earlier prognostic marker for disease progression than decline in the level of antibodies to p24. The aim of this study was to clone and express the gag derived p17 protein in soluble form in E. coli and then assess the immunoreactivity of produced recombinant p17. DNA sequence encoding p17 matrix protein was cloned from PTZ-gag53-IR vector that has the complete gag polyprotein sequence. The T7-promoter-based expression system used in this study was TOPO directional cloning strategy that expressed p17 matrix protein in fusion with thioredoxin in E. Coli. Purification of produced recombinant protein has been done using Ni-NTA nickel chelating agarose beads. Sequencing result of the cloned sequence showed that it belongs to CRF35_AD subtype of HIV-1 which is highly prevalent in Iran and Afghanistan. The immunoreactivity of produced<br />recombinant p17 to sera from infected individuals showed 93.8% sensitivity and 100% specificity.<br /><br />https://www.ijbiotech.com/article_7177_be264ec9eae6e312ef0357f8e60c642f.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304310420121001Periplasmic expression of Bacillus thermocatenulatus lipase in Escherichia coli in presence of different signal sequences2552627164ENAli Asghar KarkhaneDepartment of Industrial and Environmental Biotechnology at the National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.Bagher YakhchaliDepartment of Industrial and Environmental Biotechnology at the National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.0000-0003-4031-877xFerdous Rastgar JaziiDepartment of Industrial and Environmental Biotechnology at the National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.Jafar HemmatDepartment of Biotechnology, Iranian Research Organization for Science and Technology (IROST),P.O. Box 15815-3538Parvin ShariatiDepartment of Industrial and Environmental Biotechnology at the National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.Mahvash KhodabandehDepartment of Industrial and Environmental Biotechnology at the National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.Alireza ZomorodipoorDepartment of Industrial and Environmental Biotechnology at the National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.Journal Article20121001Efforts to express lipase in the periplasmic space of Escherichia coli have so far been unsuccessful and<br />most of the expressed recombinant lipases accumulate in the insoluble cell fraction. To evaluate the role of<br />native and heterologous signal peptides in translocation of the lipase across the inner membrane of E. coli,<br />the lipase gene (btl2) was cloned downstream of the native Bacillus signal peptide and also in fusion with<br />the pelB, ansB and ansB/asp signal peptides. For this purpose, four recombinant expression vectors (pYRKP.P, pYRKP.N, pYRKP. A and pYRKP.AA) were constructed and expressed in E. coli. Osmotic shock analysis showed that recombinant lipase was overexpressed as inclusion bodies in E. coli. The lipase inclusion bodies were subsequently solublized, refolded and purified using single column ion-exchange chromatography. To evaluate localization of lipase in the cell, the purified lipases were subjected to capillary isoelectric focusing and tandem mass spectrometry. Results showed that all signal peptides were able to direct the lipase from the cytoplasm into the periplasmic space of E. coli, because the periplasmic space of E. coli is not suitable for lipase folding, the translocated lipase aggregates in this space as inclusion bodies.https://www.ijbiotech.com/article_7164_cf7cdb23a04a72c97a1c08fa099988aa.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304310420121001Comparison between batch and fed-batch production of rhamnolipid by Pseudomonas aeruginosa2632697170ENMohammad Ghomi AviliDepartment of Chemical Engineering, Shahid Bahonar University of Kerman, P.O. Box 76169133, Kerman, I.R.Iran.Mohammad Hassan FazaelipoorDepartment of Chemical Engineering, Shahid Bahonar University of Kerman, P.O. Box 76169133, Kerman, I.R.Iran.Seyed Ali JafariDepartment of Chemical Engineering, Shahid Bahonar University of Kerman, P.O. Box 76169133, Kerman, I.R.Iran.Seyed Ahmad AtaeiDepartment of Chemical Engineering, Shahid Bahonar University of Kerman, P.O. Box 76169133, Kerman, I.R.Iran.Journal Article20121001This paper presents a comparison between batch and three different sets of fed batch fermentations for<br />rhamnolipid production by Pseudomonas aeruginosa. The batch run was performed with 500 ml of culture<br />medium having the initial glycerol and sodium nitrate concentrations of 30 and 8.3 g/l, respectively. For a fed<br />batch run with nitrogen source in feed, 250 ml of the nitrogen excluded culture medium was in the bioreactor<br />initially, and 250 ml culture medium containing 16.6 g/l sodium nitrate was fed to the bioreactor continuously.<br />A similar procedure was repeated for fed batch runs with carbon, and phosphorus source in feed. Statistical<br />analysis showed that fed batch runs were better than batch in term of rhamnolipid production, and among<br />the fed batch runs the maximum amount of rhamnolipid. (4.12 g Rhamnose Equivalent/l) was for the fed<br />batch run with the carbon source in feed.https://www.ijbiotech.com/article_7170_3fe58d25bcb7df9b9f758bb95e1a4f7d.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304310420121001Expression of cytochrome P450 and glutathione S-transferase in human bone marrow mesenchymal stem cells2702747187ENShahnaz EsmaeliDepartment of Clinical Biochemistery, Tarbiat Modares University, P.O. Box 14115-331, Tehran, I.R. Iran. Faculty of
Nursing and Midwifery, Tehran University of Medical Sciences, P.O. Box 1419733171, Tehran I.R. Iran.Abdolamir AllamehDepartment of Clinical Biochemistery, Tarbiat Modares University, P.O. Box 14115-331, Tehran, I.R. Iran.Mohammad Sajad Emami AleaghaDepartment of Clinical Biochemistery, Tarbiat Modares University, P.O. Box 14115-331, Tehran, I.R. Iran.Somaieh KazemnejadDepartment of Clinical Biochemistery, Tarbiat Modares University, P.O. Box 14115-331, Tehran, I.R. Iran.Masoud SoleimaniDepartment of Hematology, Tarbiat Modares University, P.O. Box 14115-331, Tehran, I.R. Iran.Journal Article20121001Currently several studies are being carried out on various properties of mesenchymal stem cells (MSCs)<br />however there are a few investigations about drug metabolizing properties of these cells. The aim of this<br />study was to measure the key factors involved in drug metabolism in human bone marrow MSCs. For this<br />purpose, cellular glutathione (GSH), glutathione Stransferase (GSTs) and cytochrome P450 class 3A4<br />(CYP3A4) were detected in these cells. Results showed that CYP3A4 and GSTA1-1 mRNA are not detectable in MSCs however mRNA specific for GSTP1-1 was considerably expressed in MSCs. GSH content and GST activity was also detected in MSCs. These data suggest that human bone marrow MSCs possess the drug metabolizing activity which may be useful in handling drugs and chemotherapeutic agents passing to the bone marrow.<br /><br />https://www.ijbiotech.com/article_7187_3669cc20e7b5983c6b9ca1d2111dcfd1.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304310420121001Optimization of secretory expression of recombinant hGM-CSF in high cell density cultivation of recombinant Escherichia coli using Taguchi statistical method2752807190ENElmira HajiniaDepartment of Chemical Engineering, Science and Research Branch, Islamic Azad University, P.O. Box 1477893855, Tehran, I.R. Iran.Seyed Safa-ali FatemiDepartment of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.Ali Asghar KarkhaneDepartment of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.Ali Akbar SafekordiDepartment of Chemical Engineering, Science and Research Branch, Islamic Azad University, P.O. Box 1477893855, Tehran, I.R. Iran.Bagher YakhchaliDepartment of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.0000-0003-4031-877xJournal Article20121001Human granulocyte macrophage colony stimulating factor (hGM-CSF) has many therapeutic applications.<br />In this study, in order to verify the purification process, the effect of carbon source, IPTG concentration and<br />post-induction time on the secretion of recombinant hGM-CSF into the culture medium by recombinant<br />Escherichia coli during high cell density cultivation were evaluated by using the Taguchi statistical method. The results indicated that glucose, 1mM IPTG and a time of 6 h post-induction, represented optimum conditions. The secreted hGM-CSF, overall volumetric productivity and purified hGM-CSF were 373 mg/l, 18 mg/l/h and 63 mg/l, respectively.https://www.ijbiotech.com/article_7190_57c232efb7cc443241a465c5edbddc02.pdf