National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304311120130101The Food Crisis and New Technologies12724410.5812/ijb.10276ENAbbas Sahebghadam LotfiNational Institute of Genetic Engineering and Biotechnology, Tehran, IR Iran.Journal Article20150114<span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 9pt;">The present crisis in global economy, the issue of cli <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 9pt;">mate change and the fast-growing world population, <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 9pt;">leading to increased demand of food, are signifiant <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 9pt;">factors reinforcing moves towards inclusive technology <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 9pt;">developments. Increasing population and consumption <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 9pt;">growth will lead to the increasing global demand for <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 9pt;">food, as well. The growing competition for land, water, <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 9pt;">and energy affct food production capability, entailing <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 9pt;">the necessity for reducing the effcts of overexploita <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 9pt;">tion of food system on the environment (1). </span></span></span></span></span></span></span></span></span></span>https://www.ijbiotech.com/article_7244_1d90980dce12373b40ee3baa80c50462.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304311120130101PreRkTAG: Prediction of RNA Knotted Structures Using Tree Adjoining Grammars313723810.5812/ijb.9213ENHesamedin Torabi DashtiDepartment of Mathematics, University of Wisconsin, Madison, USAFatemeh Zare-MirakabadDepartment of Computer Science, Faculty of Mathematics and Computer Science, Amirkabir University of Technology, Tehran, IR IranNima AghaeepourDepartment of Computer Science, University of British Columbia, Vancouver, CanadaHayedeh AhrabianDepartment of Computer Science , School of Mathematics, Statistics, and Computer Science, University of Tehran, Tehran, IR IranAbbas Nowzari-DaliniDepartment of Computer Science , School of Mathematics, Statistics, and Computer Science, University of Tehran, Tehran, IR IranJournal Article20130114<span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-BoldItalic; color: #231f20; font-size: 8pt;"><em>Background: </em><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">RNA molecules play many important regulatory, catalytic and structural <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">roles in the cell, and RNA secondary structure prediction with pseudoknots is one the <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">most important problems in biology. An RNA pseudoknot is an element of the RNA sec <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">ondary structure in which bases of a single-stranded loop pair with complementary bases <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">outside the loop. Modeling these nested structures (pseudoknots) causes numerous com <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">putational diffilties and so it has been generally neglected in RNA structure prediction <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">algorithms. <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-BoldItalic; color: #231f20; font-size: 8pt;"><em>Objectives: </em><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">In this study, we present a new heuristic algorithm for the Prediction of RNA <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">Knotted structures using Tree Adjoining Grammars (named PreRKTAG). <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-BoldItalic; color: #231f20; font-size: 8pt;"><em>Materials and Methods: </em><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">For a given RNA sequence, PreRKTAG uses a genetic algorithm on <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">tree adjoining grammars to propose a structure with minimum thermodynamic energy. <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">The genetic algorithm employs a subclass of tree adjoining grammars as individuals by <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">which the secondary structure of RNAs are modeled. Upon the tree adjoining grammars, <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">new crossover and mutation operations were designed.The finess function is defied ac <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">cording to the RNA thermodynamic energy function, which causes the algorithm conver <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">gence to be a stable structure. <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-BoldItalic; color: #231f20; font-size: 8pt;"><em>Results: </em><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">The applicability of our algorithm is demonstrated by comparing its iresults with <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">three well-known RNA secondary structure prediction algorithms that support crossed <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">structures. <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-BoldItalic; color: #231f20; font-size: 8pt;"><em>Conclusions: </em><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">We performed our comparison on a set of RNA sequences from the RNAseP <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">database, where the outcomes show effiency and practicality of the proposed algorithm.</span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span><br style="line-height: normal; widows: 2; text-transform: none; font-variant: normal; font-style: normal; text-indent: 0px; white-space: normal; orphans: 2; letter-spacing: normal; font-weight: normal; word-spacing: 0px; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px;" /><br class="Apple-interchange-newline" /></span></span></span></span></span></span>https://www.ijbiotech.com/article_7238_09cd11be110ad647909e94d340754f71.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304311120130101Preparation of Antibody Against Immunodominant Membrane Protein (IMP) of Candidatus Phytoplasma aurantifolia1421720510.5812/ijb.9305ENFatemeh ShahryariDepartment of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran, IR IranMasoud Shams-BakhshDepartment of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran, IR IranMohammad Reza SafarnejadDepartment of Microbial Biotechnology and Biosafety, Agricultural Biotechnology Research Institute of Iran, Karaj, IR Iran
Department of Plant Viruses, Iranian Research Institute of Plant Protection, Tehran, IR IranNaser SafaieDepartment of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran, IR IranSaeed Ataei KachoieeRazi Vaccine and Serum Research Institute, Tehran, IR IranJournal Article20130114<span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-BoldItalic; color: #231f20; font-size: 8pt;"><em>Background: </em><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">The witches’ broom disease of lime caused by <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Italic; color: #231f20; font-size: 8pt;"><em>Candidatus </em><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">Phytoplasma au<span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">rantifolia, is the most devastating disease of acidian lime in the southern parts of Iran. </span></span></span></span></span>
<span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-BoldItalic; color: #231f20; font-size: 8pt;"><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;"><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Italic; color: #231f20; font-size: 8pt;"><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;"><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;"><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-BoldItalic; color: #231f20; font-size: 8pt;"><em>Objectives: </em><span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">At present, no effient method has been developed for controlling the dis<span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">ease, therefore quarantine approaches such as early detection and subsequent eradica<span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">tion of infected trees is very important. Toward this aim, developing a reliable and sensi<span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">tive detection method would be the fist step to prevent transportation of infected plant <span style="font-variant: normal; font-style: normal; font-family: CapitoliumNews-Regular; color: #231f20; font-size: 8pt;">materials to other places.</span></span></span></span></span></span></span></span></span></span></span>
<em>Background: </em>The witches’ broom disease of lime caused by <em>Candidatus </em>Phytoplasma aurantifolia, is the most devastating disease of acidian lime in the southern parts of Iran.
<em>Objectives: </em>At present, no efficient method has been developed for controlling the disease, therefore quarantine approaches such as early detection and subsequent eradication of infected trees is very important. Toward this aim, developing a reliable and sensitive detection method would be the first step to prevent transportation of infected plant materials to other places.
<em>Materials and Methods: </em>In this study, Immunodominant membrane protein (IMP) of the pathogen was selected as a target for detection and preparation of polyclonal antibody. The IMP is the major protein present on the surface of phytoplasma cells. For this purpose, the DNA region encoding IMP gene was isolated and cloned into pET28a bacterial expression vector. The recombinant protein was expressed in a large scale in <em>Escherichia coli.</em> Purification was performed under native conditions and the purity and integrity of produced recombinant protein were confirmed by western immuno blot analysis using anti His-tag and anti-IMP polyclonal antibodies. The purified recombinant IMP was used for immunization of rabbit. Purification of immunoglobulin was performed by affinity chromatography using protein A column. The purified immunoglobulin was conjugated with the alkaline phosphatase enzyme.
<em>Results:</em> The purified antibodies and conjugates were applied for efficient detection of infected plants in double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and dot immunosorbent assay (DIBA).
<em>Conclusions:</em> These antibodies were proven to be very powerful tools to detect the <em>Candidatus</em> Phytoplasma aurantifolia in plants.
https://www.ijbiotech.com/article_7205_4bc5132eef096b5a3fc00a1415e5817f.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304311120130101Discovering Domains Mediating Protein Interactions2231722910.5812/ijb.9309ENChangiz EslahchiDepartment of Computer Sciences, Shahid Beheshti University GC, Tehran, IR Iran0000000000000000Journal Article20130114<em>Background: </em>Protein-protein interactions do not provide any direct information regarding the domains within the proteins that mediate the interactions. The majority of proteins are multi domain proteins and the interaction between them is often defined by the pairs of their domains. Most of the former studies focus only on interacting domain pairs. However they do not consider the interactions that require the presence of a third domain. <em>Objectives:</em> In this manuscript, we define the concept of necessary and sufficient triplets of domains and mediator domain. <em>Materials and Methods:</em> We approximate these conditions by pragmatic statistical definitions on a set of gold-standard interacting protein pairs and a set of gold-standard non-interacting protein pairs. <em>Results:</em> In this paper we introduce a new method for the prediction of the interaction between two domains using third domains as a mediator.we show that the mediator domain has an effective role in the interaction between proteins. <em>Conclusions:</em> By using these concepts, we introduce a method for the prediction of the interaction between two domains. Subsequently by evaluating the performance of our method on the yeast protein interactions data set, we show that the mediator domain has an effective role in the interaction between proteins.https://www.ijbiotech.com/article_7229_e4544e06457b625313c433c34491c09c.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304311120130101Characterization and Potentials of Indigenous Oil-Degrading Bacteria Inhabiting the Rhizosphere of Wild Oat (Avena Fatua L.) in South West of Iran.3240719710.5812/ijb.9334ENSaeideh RajaeiSoil Science Department, Faculty of Agriculture, Shahrekord University, Shahrekord, IR IranSeyed Mahdi SeyediDepartment of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, IR IranFayez RaiesiSoil Science Department, Faculty of Agriculture, Shahrekord University, Shahrekord, IR IranBehrouz ShiranDepartment of Agronomy and Plant Breeding, Faculty of Agriculture, Shahrekord University, , Shahrekord, IR Iran0000-0001-9997-4718Jamshid RahebDepartment of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, IR IranJournal Article20130114<em>Background:</em> The contamination of ecosystem with petroleum and its derivatives is considered as one of the most crucial environmental threat in Iran. Application of microorganisms has been demonstrated as an appropriate and more practical alternative to clean-up petroleum hydrocarbons in the contaminated environments. <em>Objectives:</em> The objectives of this study were isolating rhizosphere-inhabiting indigenous oil-degrading bacteria in wild oat grown in petroleum-polluted areas and <em>in vitro </em>evaluating the efficiency of oil biodegradability by microbial isolates. <em>Materials and Methods:</em> Bacteria were isolated from rhizosphere of wild oat grown on contaminated sites in Khuzestan and were identified based on 16S rRNA gene sequencing. The catabolic genes were detected using PCR and hybridization analysis. Hydrocarbon degradation in liquid culture was evaluated by gas chromatography-mass spectrometry (GC-MS). <em>Results:</em> 23 indigenous oil-degrading bacterial strains were isolated from the wild oat rhizosphere, grown in severely oil contaminated soil in Khuzestan. Based on 16S rRNA gene sequence analyses, isolated strains were classified to Genera <em>Acinetobacter, Pseudomonas, Enterobacter, Stenotrophomonas, Bacillus, Achromobacter, Ochrobactrum, Paenibacillus, Microbacterium, Curtobacterium </em>and <em>Sphingobacterium.</em> Catabolic genes <em>alkM, alkB </em>and <em>xylE</em>, responsible for biodegradation of the alkanes and aromatic petrochemical compounds were detected in bacterial community inhabiting rhizosphere of the wild oat. The GC-MS analysis indicated that consortium of these bacteria was capable of reducing crude petroleum in the liquid culture by 40.5%, after 10 days. The results of the present study revealed the adaptability of microbes to the rhizospheric area and subsequently their great potential to be exploited for cleaning up hydrocarbon contaminated sites. <em>Conclusions:</em> This study might be an important step towards the development of a phytoremediation strategy in the South of Iran.https://www.ijbiotech.com/article_7197_befc9cd5a5187954de33d093ad06bf95.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304311120130101The Effect of Magnetic Fe3O4 Nanoparticles on the Growth of Genetically Manipulated Bacterium, Pseudomonas aeruginosa (PTSOX4)4146722510.5812/ijb.9302ENMohammad Esmaeel KafayatiEngineering Department, Tehran University, Tehran, IR IranJamshid RahebNational Institute of Genetic Engineering and Biotechnology, Tehran, IR IranMahmoud Torabi AngaziEngineering Department, Tehran University, Tehran, IR IranShahrokh AlizadehMicrobiology Department, Azad University of Karaj, Karaj, IR IranHassan BardaniaNational Institute of Genetic Engineering and Biotechnology, Tehran, IR Iran0000-0001-8799-173XJournal Article20130114<em>Background: </em>Magnetite (Fe3O4) nanoparticles are currently one of the important and acceptable magnetic nanoparticles for biomedical applications. To use magnetite nanoparticles for bacteria cell separation, the surface of nanoparticles would be modified for immobilizing of nanoparticles on the surface of bacteria. Functionalization of magnetite nanoparticles is performed by different surfactants such as glycine or oleic acid to attach on the bacteria cell surface simultaneously. The magnetic nanoparticles have very low toxicity on the living cells. There are some studies on evaluating the toxicity of magnetite nanoparticles on eukaryote cells, which their results showed negligible toxicity in eukaryote cells of the modified magnetite nanoparticles with different surfactants. But the toxicity of magnetite nanoparticles on bacteria cells is not reported. <em>Objectives: </em>in this study, the effect of the magnetic nanoparticles iron oxide (Fe3O4) on the growth rate of the genetically engineered <em>Pseudomonas aeruginosa </em>(PTSOX4) cells in different media with different magnetic nanoparticles concentration have been investigated. <em>Materials and Methods: </em>In this study, the genetically manipulated bacterial cells, <em>Pseudomonas aeruginosa</em> (PTSOX4), were coated with magnetic Fe3O4 nanoparticles to evaluate the toxicity effect of these nanoparticles on the growth rate of this strain in Laurial Bertany (LB) and Basal Salt media (BSM) separately. In addition the minimal inhibitory concentration (MIC) and the minimal bactericidal concentration (MBC) tests of these nanoparticles were examined. <em>Results: </em>A low concentration of nanoparticles has little toxicity effect on the cell growth in this bacterium. Maximal level of the growth obtained in the late stationary phase, using a concentration of 500 ppm or more of Fe3O4 nanoparticles, but a high concentration of these nanoparticles, more than 1000 PPM, resulted in reducing the cell growth rate. However, there was not a considerable lethal effect on the cell viability. Moreover, using a high nanoparticle concentration leads to a high level of bacterial cell coating due to more contact of the nanoparticles to bacterial cell surface. <em>Conclusions:</em> It is concluded that magnetite nanoparticles have negligible toxicity on the living bacteria cells and they are so applicable in different parts of biotechnology fields.https://www.ijbiotech.com/article_7225_27d3b863fafabe395d9f5a6899afc0a2.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304311120130101Codon Optimization, Cloning and Expression of the Human Leukemia Inhibitory Factor (hLIF) in E. coli4753719510.5812/ijb.9229ENParichehr DarabiDepartment of Genetics, Shahid Chamran University, Ahvaz, IR IranHamid GalehdariDepartment of Genetics, Shahid Chamran University, Ahvaz, IR IranSaeed Reza KhatamiDepartment of Genetics, Shahid Chamran University, Ahvaz, IR IranNahid ShahbazianDepartment of Gynecology, Ahvaz Jundishapur University of medical School, Ahvaz, IR IranMohammad ShafeeiDepartment of Genetics, Shahid Chamran University, Ahvaz, IR IranAmir JalaliResearch Center of Toxicology, Ahvaz Jundishapur University of Medical School, Ahvaz, IR IranAli KhodadadiCancer Research Center, Ahvaz Jundishapur University of Medical School, Ahvaz, IR IranJournal Article20130114<em>Background:</em> Leukemia inhibitor factor (LIF) is a very important pleiotropic cytokine which belongs to interleukin-6 (IL-6) family. LIF exerts multiple effects on different types of cells and tissues with numerous regulatory effects<em> in vivo </em>and<em> in vitro</em>. It is a lymphoid factor, which performs a number of activities including cholinergic neuron differentiation, control of stem cell pluripotency, bone and fat metabolism, and is important for embryo implantation and promoting megakaryocytes production in vivo. Human LIF is a potential therapeutic candidate for some diseases such as multiplesclerosis (MS). <em>Objectives: </em>Because of aforementioned applications of the LIF protein in biological systems, the LIF gene has been cloned in various species. In this study a useful novel method was used to clone an optimized LIF sequence. <em>Materials and Methods:</em> In this study, the optimized cDNA form of human leukemia inhibitory factorwas cloned into the pET-28a (+) expression vector under control of T7lacpromoter using BamHI and XbaI restriction enzymes. The recombinant vector was transformed into <em>Escherichia coli</em> strain BL21 (DE3). The cloned <em>hLIF </em>cDNA was expressed as a fusion protein with His-tag. Cloning of <em>hLIF</em> cDNA was confirmed by digestion and DNA sequencing. Appropriate expression of recombinant hLIF was examined by SDS-PAGE after induction with isopropylthio-β-galactoside (IPTG). <em>Results: </em>The results confirmed the expression of the 19.7 kDa rhLIF protein in the bacterial expression system. <em>Conclusions: </em>We could show that codon optimization might increase the production of recombinant hLIF in the <em>E. coli. </em>This result is useful in similar cases, in which the level of expressed gene is critical.https://www.ijbiotech.com/article_7195_ee0ae53881bbaa801b146bc83f5edd2c.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304311120130101Identification of a Specific Pseudo attP Site for Phage phiC3 Integrase in the Genome of Chinese Hamster in CHO-K1 Cell Line548720310.5812/ijb.9301ENMohammad Hadi SekhavatiDepartment of Animal Science, Ferdowsi University of Mashhad, Mashhad, IR IranKianoush DormianiDepartment of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, IR IranKamran GhaediDepartment of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, IR Iran
and
Biology Department, School of Sciences, University of Isfahan, Isfahan, IR IranYahya KhazaieDepartment of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, IR IranMorteza HosseiniDepartment of Reproductive Biotechnology, Reproductive Biomedicine Center, Royan Institute for Biotechnology, Isfahan, IR IranMojtaba TahmoorespurDepartment of Animal Science, Ferdowsi University of Mashhad, Mashhad, IR IranMohammad Reza NassiriDepartment of Animal Science, Ferdowsi University of Mashhad, Mashhad, IR IranMahboubeh ForuzanfarDepartment of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, IR IranMohammad Hossein Nasr EsfahaniDepartment of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, IR IranJournal Article20130114<em>Background:</em> PhiC31 integrase is a DNA site-specific recombinase integrates DNA into the chromosomes between the two sites of <em>attB</em> and <em>attP</em>. Several pseudo <em>attPs </em>have been identified in mammalian genomes with critical features for long-term expression of transgene. In this manuscript, we report a novel intrinsic pseudo <em>attP </em>site named CHOL1 in the Chinese hamster genome implementing an inverse Polymerase Chain Reaction (IPCR).
<em>Objectives:</em>Identification of pseudo <em>attP</em> site(s) of Tenecteplase cDNA integration in the genome of stable transformed CHO cell line.
<em>Materials and Methods: </em>First, genome was extracted from a stable transformed CHO cell line expressing Tenecteplase. By creating of minicircle DNA in the last step, sequencing was performed.
<em>Results: </em>We obtained one band. BLAST analysis of the respective sequence of inverse PCR band identified a pseudo <em>attP</em> site.
<em>Conclusions:</em> Data demonstrated that the phiC31 integrase provides a suitable insertion site in the genome to express the gene of interest.https://www.ijbiotech.com/article_7203_a1f532c53ecb5ccc5932e4281ec5db96.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304311120130101Heterologous Expression of Bovine Prochymosin in Pichia pastoris GS1155963720910.5812/ijb.9228ENFatemeh RamezaniNational Institute of Genetic Engineering and Biotechnology, Tehran, IR Iran
and
Departments of Biology, Faculty of Microbiology, Alzahra University, Tehran, IR IranSara Sadr Mohammad BeigiNational Institute of Genetic Engineering and Biotechnology, Tehran, IR Iran
and
Departments of Biology, Faculty of Microbiology, Alzahra University, Tehran, IR IranGholamreza AhmadianNational Institute of Genetic Engineering and Biotechnology, Tehran, IR IranMohammadreza SoudiDepartments of Biology, Faculty of Microbiology, Alzahra University, Tehran, IR IranSoheila GhandiliNational Institute of Genetic Engineering and Biotechnology, Tehran, IR IranJournal Article20130114<em>Objectives:</em> In present research we evaluate the expression of this critical enzyme in a eukaryotic system for future use in cheese industry.
<em>Materials and Methods:</em> We have cloned bovine prochymosin gene in methylotrophic yeast, <em>P. pastoris, </em>using pPIC9K as an expression vector. The recombinant plasmid was transformed into the host by electroporation, and it was expressed in optimum conditions (temperature 29oC, 200 rpm, 2% methanol for induction, and 5 days of incubation). Transcription and expression of the recombinant prochymosin was evaluated by the reverse transcription polymerase chain reaction (RT-PCR), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis as well as western blotting and enzyme-linked immunosorbent assay (ELISA).
<em>Results:</em> In optimum conditions, only a low level of this heterologous protein was detected using ELISA method and subsequently confirmed by RT-PCR.
<em>Conclusions: </em>Since it has been reported that <em>P. pastoris </em>is an appropriate host for the expression of recombinant proteins, a low level of expression of prochymosin in this host should be explored in our future research.https://www.ijbiotech.com/article_7209_82c94e4a06a1261f2dfab9dd4972f75c.pdf