National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304312220140401Expression, Purification and Characterization of Human Recombinant Galectin 3 in Pichia pastoris38725710.5812/ijb.17330ENPraveen Kumar VemuriDepartment of Biotechnology, K L University, Guntur District, Andhra Pradesh, IndiaSuryanarayana VeeravalliDepartment of Applied Biology, Jigjiga Univeristy, EthiopiaJournal Article20140101Background: Over the past century, the areas of genomics, proteomics and lipids have captured the attention of investigators worldwide. Carbohydrates, have recently received increased attention through the expanding field of glycobiology; probably because they are very complex and not encoded in the genome. Objectives: The purpose of this study was to express and purify recombinant human galectin 3via the <em>Pichiapastoris</em> expression system. Materials and Methods:cDNA of human galectin 3 gene was amplified with specific primers and cloned into a pcDNA3.1 vector with His-tag for easier purification using Ni2andchromatography. Furthermore, galectin 3was purified to homogeneity and confirmed using SDS-PAGE and western blotting. Results:The protein band corresponding to 29 kDa was excised from the gels, digested with trypsin and processed for mass spectrometric analysis by Matrix Assisted Laser Desorption/Ionization- Time of Flight Mass Spectroscopy (MALDI-TOF MS), using a Reflex III instrument. Conclusions:Tryptic digest analysis clearly revealed that the purified protein was indeed galectin 3. Similarly, the biological activity of recombinant galectin 3 was confirmed using the hemagglutination inhibition assay.National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304312220140401Cloning and Expression of the Variable Regions of Anti-EGFR Monoclonal Antibody in E. coli for Production of a Single Chain Antibody914725110.5812/ijb.17522ENFarzaneh JalalypourBiotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, I.R. IRAN
Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, I.R. IRANSafar FarajniaImmunology Research Center, Tabriz University of Medical Sciences, Tabriz, I.R. IRAN0000-0002-6087-9147Fatemeh MahmoudiDepartment of Cellular and Molecular Biology, Azarbaijan Shahid Madani University, Tabriz, I.R. IRANBehzad BaradaranImmunology Research Center, Tabriz University of Medical Sciences, Tabriz, I.R. IRANDavoud FarajzadehDepartment of Cellular and Molecular Biology, Azarbaijan Shahid Madani University, Tabriz, I.R. IRAN0000-0002-9267-4277Leila RahbarniaDrug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, I.R. IRANJafar MajidiImmunology Research Center, Tabriz University of Medical Sciences, Tabriz, I.R. IRANJournal Article20140101Background:Epidermal growth factor receptor (EGFR) overexpression is a characteristic of several malignancies and could be considered as an excellent target for designing specific inhibitors such as anti-EGFR monoclonal antibodies for cancer therapy. Drawbacks exerted by large sizes of full-length antibodies have lead to the development of single chain antibodies, which benefit from having smaller sizes and short circulation half-lives. Objectives:The aim of this study was cloning, expression and purification of variable regions of anti-EGFR monoclonal antibody in <em>E. coli</em> for production of single chain antibodies. Materials and Methods:The RNA, extracted from the C225 hybridoma cells, was reverse transcribed into cDNA and used for PCR amplification of genes encoding light and heavy chains from the variable regions. The PCR products were cloned and expressed in <em>E. coli</em> BL21 for production of a single chain antibody. The expressed protein was analyzed by SDS-PAGE and purified by Ni-NTA affinity chromatography. The reactivity of purified C225-scFv with EGFR-expressing A431 tumor cell line was tested by western blotting and enzyme-linked immunosorbent assays. Results:The results indicated that C225-scFv was highly expressed in <em>E. coli</em> and appeared as a protein with a mass of 27 kDa in the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the induced cell lysate. Reactivity analysis of the purified C225-scFv with A431 tumor cell line by western blotting and enzyme linked immune sorbant assay (ELISA) revealed high binding affinity of the recombinant C225-scFv to the target cells. Conclusions:The results of this study indicated that C225-scFv is capable of binding to EGFR and could be considered as a useful tool for diagnosis and treatment of EGFR-overexpressing tumor cells.National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304312220140401Simultaneous Camouflage of Major and Minor Antigens on Red Blood Cell Surface With Activated mPEGs1525726010.5812/ijb.17776ENZahra GholamiFaculty of Chemical Engineering, Tarbiat Modares University, Tehran, I.R. IRANSameereh Hashemi NajafabadiFaculty of Chemical Engineering, Tarbiat Modares University, Tehran, I.R. IRANMasoud SoleimaniFaculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. IRANJournal Article20140101<span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Bold; color: #231f20; font-size: 8pt;">Background: <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">Host immune system response against blood group antigens is a major problem in blood transfusions, especially for <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">thalassemic patients. Thus, an approach was proposed coating the red blood cell (RBC) surface by polyethylene glycol. <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Bold; color: #231f20; font-size: 8pt;">Objectives: <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">This study aimed to obtain the optimal simultaneous camouflge of the major and minor antigens by activated methoxy <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">polyethylene glycol (mPEG) with succinimidyl valerate (SVA) and succinimidyl carbonate (SC), separately. <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Bold; color: #231f20; font-size: 8pt;">Materials and Methods: <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">The degree of RBC agglutination by antibodies against the major and minor blood groups was used as a <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">surrogate measurement for quantitative assessment of the effctiveness of the surface coating. Also, the RBC morphology was assessed <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">using scanning electron microscope (SEM). In addition, to evaluate the host immune system response, the PEGylated RBCs were transferred <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">between two diffrent mouse strains. <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Bold; color: #231f20; font-size: 8pt;">Results: <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">Statistical analysis of the results demonstrated that the optimal reaction conditions for simultaneous coating of the antigens <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">by mPEG-SVA and mPEG-SC are as mPEG<span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 7pt;">20 <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">in the polymer mixture, 91.2 and 90.0%, and polymer concentration, 17.21 and 19.80 mg.mL-1, <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">respectively. However, according to the SEM results, the maximum polymer concentration of 14.5 mg.mL-1 was suggested as the best <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">condition for mPEG-SVA modifid human RBCs. <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Bold; color: #231f20; font-size: 8pt;">Conclusions: <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">It is concluded that the membrane PEGylation camouflges the blood group antigens. This effct is observed signifiantly <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">for non-ABO/Rh(D) antigens. Also, it is found that the mPEG-SVA provide better coverage than mPEG-SC. The results of in vivo analysis <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">showed that the immune reactions against PEGylated RBCs were considerably reduced, so that the levels of the relevant biochemical <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">parameters in serum were similar to those of the normal hosts 24 hours after transfusion. </span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span>National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304312220140401Enhancement of Trichoderma Harzianum Activity Against Sclerotinia sclerotiorum by Overexpression of Chit422631725310.5812/ijb.13869ENMojegan KowsariNational Institute of Genetic Engineering and Biotechnology, Tehran, I.R. IRAN
and
Agricultural Biotechnology Research Institute of Iran, Karaj, I.R. IRANMohammad Reza ZamaniNational Institute of Genetic Engineering and Biotechnology, Tehran, I.R. IRANMostafa MotallebiNational Institute of Genetic Engineering and Biotechnology, Tehran, I.R. IRANJournal Article20130727 <span style="color: windowtext;">Backgoround: </span><span style="color: windowtext;">Plant diseases, caused by a wide range of phytopathogenic fungi, could be managed using of <em>Trichoderma</em> sp, as a biocontrol agent. Cell wall degrading enzymes like chitinase from <em>T. harzianum</em> are important means for fungal pathogen inhibition. Overexpression of these chitinase enzymes can improve the antagonistic potential of <em>Trichoderma</em> sp. strains.</span> <span style="color: windowtext;">Objectives: </span><span style="color: windowtext;">This study aimed to produce a new enhanced biocontrol system of Trichoderma harzianum, overexpressing <em>chit42</em> gene. The improved <em>T. harzianum</em> could be an appropriate biocontrol agent for controlling the stem rot disease of canola caused by <em>Sclerotinia sclerotiorum</em>.</span> <span style="color: windowtext;">Materials and Methods: </span><em><span style="color: windowtext;">T. harzianum</span></em><span style="color: windowtext;"> protoplast cotransformation was carried out by pLMRS3-Chit42 and p3SR2 plasmids. The transformants were selected based on their growth on colloidal chitin containing medium. The improvement of transformants was investigated by quantification of mRNA using real-time quantitative polymerase chain reaction (RT-PCR) and measurement of chitinase activity in the medium containing colloidal chitin as the carbon source. Furthermore, the antagonistic activity of transformants against <em>S. sclerotiorum</em> was assessed by dual culture experiments. </span> <span style="color: windowtext;">Results: </span><span style="color: windowtext;">The overexpressing transformants of <em>Chit42</em> displayed higher levels of chitinase activity to inhibit <em>S. sclerotiorum</em> growth compared with the wild type. The results indicated that the value of the chitinase activity (126.42 + 0.07 U/mL) of <em>Chit42-9</em> increased 64.17 fold. Transcriptomic analysis demonstrated that <em>Chit42-9</em> transformant expressed 5.2 fold of <em>Chit42</em> transcript as compared with the parent strain. Biocontrol inhibition of this transformant was 4.98-fold more compared with the non-transformant type.</span> <span style="color: windowtext;">Conclusion: </span><span style="color: windowtext;">The improved <em>Chit42-9</em> transformant can be used for biocontrolling S. sclerotiorum, cause of stem rot disease in canola.</span>National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304312220140401Molecular Characterization and Antimicrobial Resistance of Uropathogenic Escherichia coli3240724610.5812/ijb.16833ENFatemeh MashayekhiSchool of Nursing and Midwifery, Jiroft University of Medical Sciences, Jiroft, I.R. IRANMandana MoghnyDepartment of Clinical Pathology, Shahrekod University of Medical Science, Shahrekord, I.R. IRANMotahare FaramarzpoorSchool of Nursing and Midwifery, Jiroft University of Medical Sciences, Jiroft, I.R. IRANEmad YahaghiBaqiyatallah University of Medical Sciences, Tehran, I.R. IRANEbrahim Khodaverdi DarianYoung Researchers and Elite Club, Karaj Branch, Islamic Azad University, Karaj, I.R. IRANVahideh TarhrizDepartment of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences,Tabriz, I.R. IRANBanafsheh DormaneshDepartment of Pediatric Nephrology, AJA University of Medical Sciences, Tehran, I.R. IRANJournal Article20131212Background:Urinary Tract Infections (UTIs) are the most common infectious diseases in childhood. The Uropathogenic <em>Escherichia coli</em> (UPEC) strains account for as much as 80% of UTIs. Objective:From a clinical perspective, it is important to know which virulence factors and antibiotic resistance properties are present in UPEC strains in pediatrics. Therefore, this study was carried out to investigate the molecular characterization and antimicrobial resistance of UPEC strains isolated from hospitalized patients in pediatric ward of Baqiyatallah Hospital in Tehran. Patients and Methods:One hundred and twenty-one urine specimens were collected from the patients infected with UTIs (51 boys and 70 girls). The urine samples were cultured immediately, and those with <em>E. coli</em>-positive were analyzed for the presence of antibiotic resistance genes and bacterial virulence factors using Polymerase Chain Reaction (PCR). Also, antimicrobial susceptibility testing was performed using disk diffusion methodology with Mueller–Hinton agar according to the instruction of Clinical Laboratory and Standard Institute. Results:Nineteen out of 51 (37.25%) urine samples from boys and 47 out of 70 (67.14%) urine samples from girls harbored <em>E. coli</em>. A significant difference was found between the frequency of UPEC strains in boys and girls (P <.05). High resistance levels to tetracycline (69.6%), ampicillin (69.6%) and norfloxacin (63.6%) were also observed. Totally, 1.66% of tested strains were resistant to more than 8 antibiotics. The incidence of genes encoding resistance against gentamicin (<em>aac</em> <em>(3)-IV</em>), sulfonamide (<em>sul1</em>), beta-lactams (<em>blaSHV</em> and <em>CITM)</em>, tetracycline (<em>tetA</em> and <em>tetB</em>), trimethoprim (<em>dfrA1</em>), and quinolones (<em>qnr</em>) were 25.7%, 22.7%, 83.2%, 71.1%, 19.6% and 21.2%, respectively. The most commonly detected virulence factors were <em>fim </em>(71.2%), <em>set-1</em> (66.6%), <em>iha</em> (62.1), <em>papG</em>I (59%), <em>usp</em> (56%) and <em>sen</em> (22.7%). Conclusion:Resistant strains of uropathogenic <em>E. coli </em>had the lower incidence of uropathogenic virulence factors. We suggested prescription of imipenem and amikacin to treat pediatric patients infected with UTIs.National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304312220140401Molecular Phylogeny of the Genus Lathyrus (Fabaceae-Fabeae) Based on cpDNA matK Sequence in Iran4148725410.5812/ijb.10315ENRoghayeh OskoueiyanDepartment of Biology, Ayatollah Amoli Branch, Islamic Azad University, Amol, I.R. IRANShahrokh Kazempour OsalooDepartment of Plant Biology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, I.R. IRANAtefeh AmirahmadiDepartment of Biology, Damghan University, Damghan, I.R. IRANJournal Article20130116<span style="color: windowtext;">Background<span lang="AR-SA" dir="rtl">: </span></span><span style="color: windowtext;">More than 60 species of the genus <em>Lathyrus</em> are distributed in Southwest Asia. It is the second largest genus of the tribe Fabeae, after <em>Vicia</em>, in the region (and in Iran with 23 species). In the regional Flora (Flora of Turkey<em>, </em>FloraIranicaand flora of Iran), the genus has been divided into 9-10 sections. Here we analyzed the phylogeny of <em>Lathyrus</em> and its relationship with <em>Pisum</em> based on plastid gene matK sequences.</span> <span style="color: windowtext;">Objectives:</span><span style="color: windowtext;">The present study utilized several approaches including maximum parsimony, Bayesian and maximum likelihood methods to evaluate the monophyly and relationship within the genus <em>Lathyrus</em>, both at the sectional level and species level, mainly based on the taxa growing in Iran.</span> <span style="color: windowtext;">Materials and Methods:</span><span style="color: windowtext;">A total of 52 accessions, representing 38 species of <em>Lathyrus</em>, three species of <em>Pisum</em> and four species of <em>Vicia</em> and <em>Lens</em> as out-groups, were analyzed for reconstructing the phylogenetic relationship using chloroplast gene matK sequences. Maximum parsimony, Bayesian and maximum likelihood methods were used to construct phylogenetic trees.</span> <span style="color: windowtext;">Results:</span><span style="color: windowtext;">The present study indicated that <em>Pisum</em> was nested among <em>Lathyrus</em> species. Two members of the <em>Lathyrus</em> section, <em>Clymenum</em> (<em>Lathyrus ochrus</em> and <em>L. Clymenum</em>) with <em>Pisum,</em> formed a weakly supported clade as sister to the larger polytomy comprising the remainder of the <em>Lathyrus</em> species. Several sections of <em>Lathyrus</em> including <em>Lathyrostylis</em>, <em>Lathyrus</em> and <em>Clymenum</em> were monophyletic. <em>Lathyrus roseus</em> (of the monotypic section <em>Orobon</em>) were nested among the members of section <em>Lathyrus</em>. The newly taxon described species <em>L. alamutensis</em>, endemic to Iran, were nested among other species of <em>Lathyrostylis</em>. <em>Linearicarpus</em>, <em>Orobus</em> and <em>Pratensis</em> were not monophyletic sections. <em>Pratensis</em> and the monotypic section <em>Aphaca</em> were the closest taxa. In our analysis, <em>L. Pratensis</em> formed a sister group relationship with the <em>Aphaca</em> clade, not its own section.</span> <span style="color: windowtext;">Conclusions:</span><span style="color: windowtext;">Shimodaira-Hasegawa (SH) test of the matK dataset showed that all analyzed <em>Lathyrus</em> species formed their own clade and <em>Pisum</em> was sister to them. Furthermore, when we removed the two above-mentioned <em>Lathyrus</em> species, the analysis retrieved <em>Pisum,</em> as a well-supported clade being sister to the <em>Lathyrus</em> calde.</span>National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304312220140401Optimization of Dynamic Binding Capacity of Anion Exchange Chromatography Media for Recombinant Erythropoietin Purification4955725610.5812/ijb.17352ENMina SepahiRecombinant Biopharmaceutical Production Department, Research and Production Complex, Pasteur Institute of Iran, Karaj, I.R. IRANHooman KaghazianRecombinant Biopharmaceutical Production Department, Research and Production Complex, Pasteur Institute of Iran, Karaj, I.R. IRANMina Payravi SereshkehRecombinant Biopharmaceutical Production Department, Research and Production Complex, Pasteur Institute of Iran, Karaj, I.R. IRANTahereh SadeghchehQuality Control Department, Research and Production Complex, Pasteur Institute of Iran, Karaj, I.R. IRANShahin HadadianQuality Control Department, Research and Production Complex, Pasteur Institute of Iran, Karaj, I.R. IRANMohammad Reza JebeliRecombinant Biopharmaceutical Production Department, Research and Production Complex, Pasteur Institute of Iran, Karaj, I.R. IRANFereshteh YavariScience and Research Branch, Islamic Azad University, Tehran, I.R. IRANJournal Article20140103 <span style="color: windowtext;">Background:</span><span style="color: windowtext;">The dynamic binding capacity (DBC) of a chromatography matrix in protein purification is the amount of the total protein absorbed into the matrix, before occurrence of a significant break in the breakthrough curve. Optimization of the process criteria for maximum DBC avoids extra process scale-up and reduces the processing time, costs and protein loss. Taguchi method is a simple useful tool in experimental design to estimate the optimal condition with minimum experiments.</span> <span style="color: windowtext;">Objectives:</span><span style="color: windowtext;">In this research, linear flow rate, pH and protein concentration of the feed were checked according to an L9 orthogonal Taguchi array, to estimate the best conditions for maximum DBC of Q-sepharose fast flow (QSFF) resin in recombinant human erythropoietin purification process.</span> <span style="color: windowtext;">Materials and Methods:</span><span style="color: windowtext;">A crud sample containing human recombinant erythropoietin was harvested from a cell culture of Chinese hamster ovary (CHO) cell line. Desalted harvests with different total protein concentrations (30, 40 and 50 µg.mL<sup>-1</sup>) and pH values (5, 6 and 7) were loaded into a packed column of QSFFwith different linear flow rates (60, 120 and 280 cm.h<sup>-1</sup>) up to 10% of the breakthrough curve. The total protein loading to the column was checked by UV absorbance and Lowry method, and erythropoietin concentration was measured by ELISA. Analysis of variance (ANOVA) was applied to determine the optimum condition.</span> <span style="color: windowtext;">Results:</span><span style="color: windowtext;">Finally, total protein concentration of 50 µg.mL<sup>-1</sup>, pH of 5 and flow rate of 120 cm.h<sup>-1</sup>, were anticipated as the optimal process conditions with 5.85 mg.mL<sup>-1</sup>of resin as the dynamic binding capacity.</span> <span style="color: windowtext;">Conclusions:</span><span style="color: windowtext;">Experiments with anticipated optimal criteria were performed three times and no significant difference was observed (p = 0.136, and 6.06 mg/mL as the average dynamic binding capacity).</span>National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304312220140401Construction of a Minigenome Rescue System for Measles Virus, AIK-c Strain5662725510.5812/ijb.18002ENMostafa GhaderiDepartment of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. IRANFarzaneh SabahiDepartment of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. IRANMajid SadeghizadehDepartment of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. IRAN0000-0002-2497-3152Zahra KhanlariDepartment of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. IRANAzam JamaatiDepartment of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. IRANSeyed Dawood Mousavi NasabDepartment of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. IRANNasrin Majidi GarenazDepartment of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. IRANJournal Article20140205Background:In the recent decade, the reverse genetics method has been broadly used for rescue of negative-stranded RNA viruses from cDNA or viral minigenomes. This technique has been applied to study different steps in virus replication and virus-host interactions. Reverse genetics could also be implemented for design of new vaccines. The T7 RNA polymerase activity as well as virus (nucleocapsid protein) N, (phosphoprotein) P and (Large) L proteins are necessary to rescue the virus or viral minigenome. Measles virus is a negative-stranded non-segmented RNA virus. There are useful vaccine strains to prevent measles disease. Objectives:Here, we describe the construction of a new helper cell line for rescue of measles virus minigenome. The helper cell line stably expresses T7 RNA polymerase as well as measles virus N and P proteins by tricistronic mRNA. Materials and Methods:For rescue of measles virus minigenome a stable helper cell line by using tricistronic expression vector was developed which expressed T7 RNA polymerase as well as measles virus N and P proteins. To construct the tricistronic expression vector, T7 RNA polymerase gene was cloned after cytomegalovirus (CMV) promoter and measles virus N and P proteins were under control of IRES (internal ribosome entry site) sequences. Results:Our results indicated that measles virus minigenome could be rescued in this constructed helper cell line. Conclusions:Through this system, the measles virus minigenome was rescued. Further studies are necessary to improve the rescue efficiency. This may be possible by replacing the CMV promoter with the T7 promoter.National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304312220140401In Vivo Toxicity Assessment of Bovine Serum Albumin and Dimercaptosuccinic Acid Coated Fe3O4 Nanoparticles6368725810.5812/ijb.16858ENParisa HajshafiiFalavarjan Branch, Islamic Azad University, Isfahan, I.R. IRANSoheil FatahianFalavarjan Branch, Islamic Azad University, Isfahan, I.R. IRANKahin ShahanipoorFalavarjan Branch, Islamic Azad University, Isfahan, I.R. IRANJournal Article20131214Background: Recently, applications of nanoparticles in many fields of medicine have been developed, due to their specific physical and chemical properties. Therefore assessment of their toxicity specially in the in vivo condition is necessary. Objectives: The aim of this study is to evaluation the effect of Fe<sub>3</sub>O<sub>4</sub> nanoparticles coating by biocompatible compounds on their toxicity and also comparison by noncoated nanoparticles. Materials and Methods: Wetted chemical method was used in order to synthesize Fe<sub>3</sub>O<sub>4</sub> nanoparticles. The synthesized nanoparticles were coated by BSA (Bovine Serum Albumin) and DMSA (Dimercaptosuccinic Acid) and the coating interactions were investigated by FTIR. Magnetic and structure properties of Fe<sub>3</sub>O<sub>4 </sub>and coated Fe<sub>3</sub>O<sub>4 </sub>nanoparticles were evaluated by AGFM (Alternating Gradient Force Magnetometer), TEM (Transmission Electron Microscope) and XRD (X Ray Diffraction). Toxicity assessment of Fe<sub>3</sub>O<sub>4 </sub>and coated Fe<sub>3</sub>O<sub>4 </sub>nanoparticles were studied in mice by intra peritoneally injections during a month. Liver enzymes (SGPT, SGOT, ALP, and LDH) were measured 7, 15 and 30 days post injection. Result: The synthesized nanoparticles are single phase and have the spinel structure which their size distribution in the net from is around 5 to 11 nm and in the coated form is 17 to 25 nm. Some liver enzymes were changed due to the injection of both uncoated and coated nanoparticles to mice (especially in groups who received concentrations more than 100 mg per kg of mice weight). The liver enzymes changes were more considerable in the groups received DMSA or DMSA coated in comparison with the groups received BSA or BSA coated. Chemical toxicity studies showed that there is not any irreversible effect in concentrations less than 200 mg/kg for all control and treated groups. Conclusions: The results indicate that, liver enzymes were changed during 7 and 15 days post injection measurements especially in high doses (200 mg/kg). The results of 30 days post injection measurements were changed less in comparison with the control and this is indicates that there is not any irreversible effect in liver. Moreover, DMSA coated nanoparticles were more toxic in comparison with BSA coated nanoparticles.