eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2013-07-01
11
3
144
144
10.5812/ijb.12619
7215
Polyclonal Antibodies for Detection of Witchesâ Broom Disease of Lime
Thomas Böldicke
1
Helmholtz Centre for Infection Research, Department of Molecular Structural Biology, Braunschweig, Germany
The witches’ broom disease of lime trees is a destructing disease caused by the bacterium Candidatus Phytoplasma aurantofolia (1). The disease is prevalent in Oman, United Arab Emirates and Iran. To restrict the spread of the infected trees, detection of the causative bacterium is crucial. Besides PCR-based methods, antibodies can be applied for detection of pathogen-infected cells. Recently Shahryari et al. demonstrated that polyclonal antibodies generated against a cell surface protein of the bacterium could be used for effient detection of phytoplasma-infected plant cells (2). In their very conclusive publication the authors describe generation and binding of polyclonal antibodies against the immunodominant membrane protein (IMP) of Candidatus Phytoplasma aurantifolia. Immunization of rabbits with recombinant antigen resulted in polyclonal antibodies with very high titer, which enabled the development of a highly sensitive, but simple ELISA assay for phytoplasma detection in infected plants. For studying plant cell-phytoplasma interaction, specifi antibodies against cell surface proteins of Candidatus Phytoplasma aurantifolia and the host cell would be useful.
https://www.ijbiotech.com/article_7215_2ccaabc8f57ab3445b94beb1fb116fdf.pdf
polyclonal antibodies
Witches’ Broom
lime
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2013-07-01
11
3
145
146
7217
Progress and Problems in Producing Nitrogen Dioxide-Philic Plants
Misa Takahashi
mtakahas@sci.hiroshima-u.ac.jp
1
Makiko Nakagawa
2
Hiromichi Morikawa
3
Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, Japan
Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, Japan
Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, Japan
The recently published articles in the Iranian Journal of Biotechnology inspired us to write this short communication on nitrogen dioxide NO2-philic plants (1). To cope with environmental pollution, we thought (still think) that development of methods that mitigate pollution is an urgent issue, and that NO2-philic plants that can grow with NO2 as the sole nitrogen source are instrumental for the implementation of mitigation of air pollution.
https://www.ijbiotech.com/article_7217_3fc0feadceecccb6ae96c59e7ff9753c.pdf
Plant
Nitrogen Dioxide
Air pollution
Arabidopsis thaliana
Nicotiana Plumbaginifolia
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2013-07-01
11
3
147
155
10.5812/ijb.11081
7234
MicroRNAs: Critical Regulators of mRNA Traffic and Translational Control with Promising Biotech and Therapeutic Applications
Sahar Ansari Basir
1
Khosrow Adeli
2
Molecular Structure and Function, Research Institute, The Hospital for Sick Children, University of Toronto, Toronto, Canada
Molecular Structure and Function, Research Institute, The Hospital for Sick Children, University of Toronto, Toronto, Canada
Context:MicroRNAs (miRNAs) are a class of short, endogenously-initiated, non-coding RNAs that post-transcriptionally control gene expression via translational repression or mRNA turnover. MiRNAs have attracted much attention in recent years as they play critical roles in gene expression and are promising tools with many biotech and therapeutic applications. The molecular mechanisms underlying the translational control of mRNAs are not fully understood but emerging evidence point to a key role for microRNAs in this process. Evidence Acquisition:In this review, we discuss the potential role of miRNAs as regulators of mRNA traffic and translational control, focusing on molecular mechanisms of miRNA-mediated control of eukaryotic mRNA stability and translational efficiency. Results:Translational control by miRNAs is often associated with silencing and repression of mRNAs via accumulation within cytoplasmic processing bodies (P-bodies), the site of mRNA storage and/or decay. Specific miRNAs can interact with the 3’UTR or 5’UTR of target mRNAs and regulate their stability as well as translational efficiency. A better understanding of these mechanisms is critical in advancing our knowledge of the role of these regulatory RNAs in modulating protein synthesis and controlling metabolic pathways in health and disease. Conclusions:The discovery of miRNAs and their important role in controlling many aspects of cell function and metabolism have led to considerable interest in biotech applications of miRNAs and their application in modulating specific gene expression. We thus highlight the growing biotech and therapeutic applications of miRNAs.
https://www.ijbiotech.com/article_7234_33f4605e212a0408a93260d9228222c1.pdf
MicroRNA
Processing Bodies
Translational Control
Metabolism
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2013-07-01
11
3
156
167
10.5812/ijb.9426
7239
Mathematical Models for Microbial Kinetics in Solid-State Fermentation: A Review
Davood Mazaheri
dmazaheri@mahallat.ac.ir
1
Seyed Abbas Shojaosadati
shoja@modares.ac.ir
2
Biotechnology Group, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, IR Iran
Biotechnology Group, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, IR Iran
Context:In this review, we discuss empirical and stoichiometric models, which have been developed recently in SSF processes and the influence of environmental conditions on the variables of these models. Additionally, new studies on modeling of product formation are also mentioned. Evidence Acquisition:Solid-state fermentation (SSF) is recognized as a cheap process for producing many valuable products like industrial enzymes and bioethanol. To develop, optimize, and scale-up this process, mathematical models are required. In this review, we collected all the papers regarding microbial growth and product formation modeling in SSF. The pros and cons of each model and confirmation with experimental data were also discussed. We discussed here the simple empirical growth kinetics models and the effect of environmental conditions on these models parameters, stoichiometric models and product formation models. Results:Simple empirical models are used widely in the kinetic modeling of SSF processes due to their simplicity and ease of use. However, more studies should be done in this field to make them more accurate, especially; the effect of environmental conditions, like temperature and moisture, on key variables of the model must be considered. Robust modeling methods, like stoichiometric models, are in their early stages in SSF processes and require more studies. Developing models in which transport phenomena models are coupled with the growth kinetics models can help better SSF bioreactor designing. On the other hand, to use SSF for producing valuable products, product formation models, which are not developed well in SSF processes, are necessary. Conclusions:To use SSF for producing valuable metabolites in large scales, more attention is required for modeling the SSF processes, especially for product formation models and using modern methods like stoichiometric models.
https://www.ijbiotech.com/article_7239_656cf0c951c3bf27fc2c3805e964e96b.pdf
Solid-State Fermentation
Mathematical Modeling
Growth Kinetics
Product Formation Model
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2013-07-01
11
3
168
174
10.5812/ijb.12468
7218
Heterologous Expression of Human IL-29 (IFN-λ1) in Iranian Lizard Leishmania
Amir Hossein Taromchi
1
Bahram Kazemi
bahram_14@yahoo.com
2
Sanaz Mahmazi
3
Mojgan Bandehpour
m.bandehpour@sbmu.ac.ir
4
Biotechnology Department, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran
Biotechnology Department, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran
Department of Genetics, Zanjan Branch, Islamic Azad University, Zanjan, IR Iran
Biotechnology Department, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran
and
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran
Background: Interferons with diffrent functions such as antiviral, antiproliferative and immunomodulatory actions are effctive medications for a number of diseases. One of these new interferons (IFNs) is Interleukin-29 (IL-29) belongs to the family of IFN-λ has antiviral activity and its potent in accompanying with IFN-α in treatment of HCV infection has been evaluated (clinical trial phase II). Recombinant IL-29 has been previously produced in multiple expression systems but in this study we cloned and expressed this protein in an Iranian Lizard leishmania (I.L.L) for the fist time. Objectives: The Main objective of this research was to evaluate expression of functional human IL-29 in domestic Lizard leishmania as an alternative eukaryotic expression system. Materials and Methods: The IL-29 expression cassette was constructed into a pLEXSY vector. The leishmania cells were transfected by electroporation. After selection of transfectants, the protein expression was evaluated at RNA and protein levels. Results: Expression cassette was successfully transfected to leishmania cells and expression of recombinant IL-29 was proved by RT-PCR and western blotting. Purifid protein showed 20% activity compared to standard protein. Enzymatic removal of N-glycan resulted to the shift of protein mobility on SDS-PAGE. Conclusions: Easy handling and culture of this eukaryotic host and mammalian cell like posttranslational modifiations are the main advantages of this expression host, but the expression yield of this protein is very low and it seems to be not economic for large-scale production.
https://www.ijbiotech.com/article_7218_e4c948b3e20046908197e28ea1a08947.pdf
Interferons
Interleukin-29
Recombinant
Leishmania
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2013-07-01
11
3
175
181
10.5812/ijb.12971
7240
Single Base Extension and Fourier-Transform Infra-Red Spectroscopy Techniques; Further Approaches in Discriminating Hazelnut-Adulterated Olive Oil
Leila Akbari
1
Zohreh Rabiei
rabiei@nigeb.ac.ir
2
Sattar Tahmasebi Enferadi
tahmasebi@nigeb.ac.ir
3
Sakineh Vanaii
4
National Institute of Genetic Engineering and Biotechnology, Tehran, IR Iran
National Institute of Genetic Engineering and Biotechnology, Tehran, IR Iran
National Institute of Genetic Engineering and Biotechnology, Tehran, IR Iran
National Institute of Genetic Engineering and Biotechnology, Tehran, IR Iran
Background:Confirmation of olive oil authenticity and particularly virgin olive oil has a great importance. Several advanced chemical and genetic analyses have been used to monitor especial components; however each has its limitations especially when detecting hazelnut-adulterated olive oil. Objectives:The objective of this research is to assess the presence of trace amount of hazelnut oil in olive oil (less than 10%) by Single Base Extension (SBE) and Fourier Transform InfraRed Spectroscopy (FTIR). Materials and Methods:The study was based on the analysis of chloroplast DNA sequences using SBE to detect Single Base Polymorphism (SNPs) in highly preserved DNA regions among species to differentiate pure and adulterated olive oil by means of two parallel tools; ABI PRISM automatic sequencer and the SNaPshot lit (APPLERA). Fourier -Transform InfraRed technique has been used for FTIR spectrum comparisons of pure olive oil and hazelnut-adulterated one as well. Results:Total DNA was extracted successfully from pure and hazelnut-adulterated olive oil, and it provided properly acceptable amplification with the primers designed on chloroplast region of both species and admixture oil of them in different ratios; 50: 50, 70: 30, and vice versa. However, for lesser than 10% hazelnut oil in olive oil only SBE analysis provided recognizable results. FTIR spectra of oil samples were assessed at frequency regions of 4000 - 700 cm-1. Eight wave numbers (3007, 1373, 1237, 1120, 1098, 1032, 965, and 722 cm-1) out of eleven differentiating ones were selected as candidate wave-numbers to distinguish pure and adulterated olive oil. Conclusions:SBE technique proved to be an effective strategy to verify olive oil authenticity, especially from hazelnut-adulterated olive oil. However, FTIR technique provided trustable results only when higher than 10% hazelnut oil is present in olive oil.
https://www.ijbiotech.com/article_7240_95284b1341d50c562dd0301bb739564c.pdf
cpDNA
FTIR
Genetic Analysis
Olive Oil Traceability
rbcL Sequence
SNPs
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2013-07-01
11
3
182
192
10.5812/ijb.9950
7198
Cloning, Expression and Characterization of Recombinant Human Fc Receptor Like 1, 2 and 4 Molecules
Mahdi Shabani
1
Azam Hemmati
2
Mahdi Zandemami
3
Jalal Khoshnoodi
4
Mahmood Jeddi-Tehrani
5
Hodjatallah Rabbani
6
Zahra Amirghofran
amirghz@sums.ac.ir
7
Fazel Shokri
fshokri@tums.ac.ir
8
Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IR Iran
and
Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, IR Iran
Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IR Iran
Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IR Iran
Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran
Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IR Iran
Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IR Iran
Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, IR Iran
Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, IR Iran
Background: The Fc receptor like (FCRL) molecules belong to the immunoglobulin (Ig) superfamily with potentially immunoregulatory function. Among the FCRL family FCRL2 and 4 are predominantly expressed on memory B cells and FCRL1 is a pan- B cell marker. To date, no ligand has been identifid for the human FCRL1, 2 and 4 molecules. Objectives: Cloning, expression, purifiation and structural analysis of the extracellular domain of human FCRL1, 2 and 4 proteins. Materials and Methods: In this study, the extracellular part of human FCRL1, 2 and 4 were subcloned into prokaryotic expression vectors pET-28b (+) and transformed into BL21-DE3 E.coli strain. Protein expression was optimized by fie adjustments such as induction time, incubation temperature and expression hosts. Recombinant FCRL proteins were purifid by metal affity chromatography using Ni-NTA resin. Purifid FCRL proteins were further characterized by SDS-PAGE and immunoblotting using His-tag and FCRL specifi polyclonal antibodies. Results: Our results demonstrated that FCRL1, 2 and 4 were successfully expressed in pET-28b (+) vector. Optimization of the expression procedure showed that IPTG induction at OD600 = 0.9 and overnight incubation at 37˚C resulted in the highest expression levels of FCRL proteins ranging from approximately 15% (FCRL1) to 25% (FCRL2 and 4) of the total bacterial lysate proteins. Conclusions: These purifid recombinant proteins are potentially a valuable tool for investigating the immunoregulatory function of FCRL molecules and the production of specifi mAbs for immunotherapeutic interventions.
https://www.ijbiotech.com/article_7198_49132f6c7406d1140cdc8c0dbe33e594.pdf
FCRL
His-tag
Polyclonal Antibody
Protein expression
Purification
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2013-07-01
11
3
193
198
10.5812/ijb.10149
7201
Protective Properties of Nontoxic Recombinant Exotoxin A (Domain I-II) Against Pseudomonas aeruginosa Infection
Asghar Tanomand
1
Shahin Najar Peerayeh
najarp_s@modares.ac.ir
2
Safar Farajnia
farajnia@gmail.com
3
Jafar Majidi
4
Department of Bacteriology, Faculty of Medical sciences, Tarbiat Modares University, Tehran, IR Iran
Department of Bacteriology, Faculty of Medical sciences, Tarbiat Modares University, Tehran, IR Iran
Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, IR Iran
Department of Immunology and Immunology Research Center, Faculty of Medical Sciences, Tabriz University of Medical Sciences, Tabriz, IR Iran
Background: Antibiotic resistance and the need for long-term treatments especially for chronic infections necessitate the development of a vaccine against Pseudomonas aeruginosa infection. Objectives: In this study, recombinant exotoxin A (domains I and II), (ExoA I-II) protein was expressed, purifid and its immunological characteristics were evaluated in a mouse model. Materials and Methods: The genomic DNA was extracted from P. aeruginosa strain PAO1. The DNA encoding for domains I and II of exotoxin A was amplifid by PCR and cloned into the pET22b expression vector. The construct was then transformed into E. coli BL21 and the protein expression was evaluated by the SDS-PAGE method. The Ni-NTA affity chromatography was used for recombinant protein purifiation. Mice were then immunized subcutaneously on day 0, 21, 42 and 72 with exotoxin A (Domains I, II). Antibody production was evaluated by the ELISA method. The immunized and control group mice were exposed to an approximate 2 × LD50 (7.5 × 107 CFU) of clinical strain of mucoid P. aeruginosa. Results: Sequencing of the cloned gene showed that the sequence of ExoA I-II gene was in accordance with ExoA I-II from P. aeruginosa PAO1. SDS-PAGE analysis indicated the expression of recombinant protein with a molecular weight of 45 KDa. Vaccination with ExoA I-II produced a signifiant amount of specifi IgG antibodies in mice. Also immunization of mice with ExoA I-II increased survival times against intra-peritoneal challenge with an approximate 7.5 × 107 CFU (2 × LD50) of clinical strain of P. aeruginosa. Conclusions: Results of this study suggested that recombinant ExoA I-II is a highly immunogenic protein which can be used as a new vaccine candidate against P. aeruginosa.
https://www.ijbiotech.com/article_7201_ba74e2fb94f8872a0218dbca9527be79.pdf
Exotoxin A
Pseudomonas aeruginosa
Recombinant Vaccine
Vaccine candidate
eng
National Institute of Genetic Engineering and Biotechnology of Iran
Iranian Journal of Biotechnology
1728-3043
2322-2921
2013-07-01
11
3
199
204
10.5812/ijb.12197
7222
CYP1B1 L432V Polymorphism and Lung Cancer Risk in the Iranian Population
Majid Motovali-Bashi
mbashi@sci.ui.ac.ir
1
Mostafa Biglari
2
Halimeh Rezaei
3
Fariba Dehghanian
fd.dehghanian@gmail.com
4
Genetics division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, IR Iran
Genetics division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, IR Iran
Genetics division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, IR Iran
Genetics division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, IR Iran
Background: Lung cancer is considered as one of the most frequent cancers worldwide, and has been the cause of more than one million mortalities each year. Exposure to tobacco smoke is the primary cause of most lung cancers, since it contains several thousand compounds, including more than 50 known carcinogens. However, a small fraction of individuals who are exposed to tobacco smoke develop lung cancer, therefore genetic factors may render some tobacco smokers more susceptible to cancer. Objectives: Genetic polymorphism in genes that encode metabolizing enzymes may be related to diffrentiated susceptibility of malignancy. CYP1B1 protein is a member of the more signifiant CYP1 subfamily enzymes, involved in environmental carcinogen metabolic activation. The most studied polymorphism in CYP1B1 gene includes 4325 C→G, resulting in an amino acid change from leucine to valine amino acid. Materials and Methods: A case-control study (included 65 lung cancer cases and 80 healthy controls) was designed based on the RFLPPCR method to estimate the possible association of this polymorphism with lung cancer susceptibility in the Iranian population. Results: Regarding the distribution of CYP1B1 L432V genotypes, there were no meaningful diffrences among controls and lung cancer patients, however among patients carrying the CC genotype, tobacco smokers had a considerable elevated risk for lung cancer compared to those who had the GG genotype. Conclusions: CYP1B1 L432V polymorphism has an important role in lung cancer risk. Therefore, further studies are recommended for investigation of other related CYP1B1 gene polymorphisms, their association with affctive genes and regulatory factors in the Iranian population.
https://www.ijbiotech.com/article_7222_3ff4dc25f1b4340fb5790a8e79059a43.pdf
Case-control study
Genetic polymorphism
Lung cancer