National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304313120150301Preparation and Characterization of Double Shell Fe3O4 Cluster@Nonporous SiO2@Mesoporous SiO2 Nanocomposite Spheres and Investigation of their In Vitro Biocompatibility110980810.15171/ijb.1068ENForough ToubiDepartment of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran
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Department of Chemical Engineering, Babol Noshirvani University of Technology, Babol, IranAbdolkhalegh DeezagiDepartment of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran0000-0001-5748-2336Gurvinder SinghDepartment of Materials Science and Engineering, Norwegian University of Science and Technology, Trondheim, NorwayMohammad Ali OghabianResearch Center for Molecular and Cellular Imaging, Tehran University of Medical Science, Tehran, IranSeyed Safa Ali FatemiDepartment of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran0000-0003-0170-8273Ayyoob ArpanaeiDepartment of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, IranJournal Article20141207Background: Multifunctional core-shell magnetic nanocomposite particles with tunable characteristics have been paid much attention for biomedical applications in recent years. A rational design and suitable preparation method must be employed to be able to exploit attractive properties of magnetic nanocomposite particles. Objectives: Herein, we report on a simple approach for the synthesis of magnetic mesoporous silica nanocomposite particles (MMSPs), consisted of a Fe3O4 cluster core, a nonporous silica shell and a second shell of the mesoporous silica of suitable sizes for biomedical applications and evaluate their cytotoxicity effects on human cancer prostate cell lines. Materials and Methods: Clusters of magnetite (Fe3O4) nanoparticles were coated by a layer of nonporous silica using Stöber method. The coating step was completed by an outer layer of mesoporous silica via template-removing method. Structural properties of MMSPs were investigated by FTIR, HR-S(T)EM, BET, XRD techniques and magnetic properties of MMSPs by VSM instrument. MTT and LDH assays were employed to study the cytotoxicity of MMSPs. Results: Obtained results revealed that decreasing the precursor concentration and the reaction time at the nonporous silica shell formation step decreases the thickness of the nonporous silica shell and consequently leads to the formation of smaller MMSPs. The as-prepared MMSPs have a desirable average size of 180±10 nm, an average pore size of 3.01 nm, a high surface area of 390.4 m2.g-1 and a large pore volume of 0.294 cm3.g-1. In addition, the MMSPs exhibited a superparamagnetic behavior and a high magnetization saturation value of 21±0.5 emu/g. Furthermore, the viability tests of DU-145 cell lines exposed to various concentrations of these particles demonstrated negligible cytotoxicity effects of the as-prepared particles. Conclusions: These results demonstrate interesting properties of MMSPs prepared in this study for biomedical applications.https://www.ijbiotech.com/article_9808_10e81563e29196adde1b911ea6e3653c.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304313120150301Stable Transformation of Saintpaulia ionantha by Particle Bombardment1116797210.15171/ijb.1037ENZahra GhorbanzadeDepartment of Biotechnology, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, IranMohammad AhmadabadiDepartment of Biotechnology, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran0000-0002-0466-7051Journal Article20141013Background: A highly efficient genetic transformation system is essential for a successful genetic manipulation of the African violet (Saintpaulia ionantha Wendl.). Objectives: Developing a particle bombardment-based genetic transformation system for the African violet. Materials and Methods: A local cultivar of the African violet from Guilan province was used for transformation experiments. The pFF19G and pBin61-Ech42 vectors were used for transient and stable transformation experiments, respectively. The PCR and RT-PCR techniques were used to verify transgene presence and transcript levels in candidate transgenic lines, respectively. Results: Using leaf explants as target tissues, we transferred an endochitinase gene cDNA into African violet. Transgenic plants were regenerated on selection medium at a reasonable frequency (in average, one stable transgenic line per shot). Molecular analysis of transgenic plants by PCR and RT-PCR techniques confirmed successful integration and expression of transgene in several independent transgenic lines. Conclusions: Our results provide an efficient stable transformation system for genetic transformation of African violet.https://www.ijbiotech.com/article_7972_d4966b4373b8a0d262b13a742103fcda.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304313120150301Functional Analysis of a Pomegranate (Punica granatum L.) MYB Transcription Factor Involved in the Regulation of Anthocyanin Biosynthesis17251055710.15171/ijb.1045ENGhazale KhaksarDepartment of Agronomy and Plant Breeding, College of Agriculture, Isfahan University of Technology, Isfahan, IranBadredin Ebrahim Sayed TabatabaeiDepartment of Biotechnology, College of Agriculture, Isfahan University of Technology, Isfahan, Iran
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Department of Molecular Plant Physiology, Utrecht University, Padualaan 8, 3584CH, Utrecht , The NetherlandsAhmad ArzaniDepartment of Agronomy and Plant Breeding, College of Agriculture, Isfahan University of Technology, Isfahan, IranCyrus GhobadiDepartment of Horticulture, College of Agriculture, Isfahan University of Technology, Isfahan, IranEsmaeil EbrahimieDepartment of Crop Production and Plant Breeding, College of Agriculture, Shiraz University, Shiraz, 73761, Iran
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Discipline of Genetics, School of Biological Science, University of Adelaide, Adelaide, 5001, SA, Australia0000-0002-4431-2861Journal Article20141023Background: Pomegranate fruit (Punica granatum L.) is a rich source of anthocyanin pigments resulting in vibrant colours and anti-oxidant contents. Although the intensity and pattern of anthocyanin biosynthesis in fruit are strongly influenced by R2R3-MYB transcription factors, little is known about the regulation and role of MYB in anthocyanin pathway of pomegranate. Objectives: The present study was conducted to elucidate the relationship between the expression of MYB transcription factor and the anthocyanin accumulation during the colour development phase of pomegranate fruits. Materials and Methods: In this work, R2R3-MYB transcription factor (PgMYB) was isolated and characterized from pomegranate skin through RACE-PCR. The expression of PgMYB gene was monitored in three distinct pomegranate accessions with distinctive skin colour and pattern by semi-quantitative RT-PCR. Results: The results indicated a strong association between skin colour in mature pomegranate fruits with the PgMYB transcripts. The highest expression level of PgMYB gene was observed in Poost Siyah Yazd (dark purple skin) throughout the ripening process. Furthermore, comparison of PgMYB amino acid sequences with those of R2R3-MYB family in grapevine, eucalyptus, peach, cacao, populus and Arabidopsis demonstrated that this protein shares high similarity (75-85% amino acid identity) with their conserved MYB domain. Computational structure prediction of PgMYB showed that the three conserved amino acids (Asn, Lys and Lys) are present in the same position of the MYB domain. Conclusions: It is speculated that PgMYB gene influences the fruit colour and could be used to improve the accumula-tion of anthocyanin pigments in the pomegranate fruit.https://www.ijbiotech.com/article_10557_90ff68581d8f58a6e8db1dd27bfd1dee.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304313120150301A simplified Protocol to Induce Callogenesis in Protoplasts of Date Palm (Phoenix dactylifera L.) Cultivars26351055810.15171/ijb.1054ENKheyreddine TitouhNational Institute of Agronomic Resaerch of Algeria, AlgeriaLakhdar KhelifiDepartment of Plant Production, LRGB- National School of Agronomy (ENSA), AlgeriaMajda SlaouiDepartment of Plant Production, LRGB- National School of Agronomy (ENSA), AlgeriaNazim BoufisNational Institute of Agronomic Resaerch of Algeria, AlgeriaAbdelkader MorsliDepartment of Plant Production, LRGB- National School of Agronomy (ENSA), AlgeriaKhadidja Hadj MoussaDepartment of Plant Production, LRGB- National School of Agronomy (ENSA), AlgeriaAbdullah MakhzoumDepartment of Biology, University of Western Ontario, London, ON CanadaJournal Article20141103Background: In Algeria, date palm is currently confronted to the Bayoud disease. Biotechnological tools such as protoplasts fusion can appear as an alternative to ensure rapid multiplication and improvement of this species. Objectives: Callogenesis induction in protoplasts isolated from embryogenic callus of three date palm cultivars. Materials and Methods: Some factors influencing the isolation and culture of protoplasts segregated from the calli of three date palm (Phoenix dactylifera L.) cultivars (Deglet Nour, Akerbouch and Degla Beida) were studied. Protoplasts of each cultivar were cultured on a semi-solid medium supplemented with various hormonal balances. Results: Maceration with an enzymatic solution containing 1.5% cellulase and 1% macerozyme R10 in the presence of 0.5 M mannitol for more than 16 h with gentle agitation allows isolation of a great number of viable protoplasts. In addition, purification of protoplasts on a cushion of 21 or 25% sucrose was effective in cell debris removal and maximum recovery. The culture of isolated protoplasts on a semi-solidified Murashige and Skoog medium, with 0.3% agarose, 2 mg. L-1 2,4-D and 0.5 mg.L-1 BAP allowed good viable protoplast maintenance as well as cell wall regeneration. After more than two months of culture, cell divisions were still occurring and microcalli became visible to the naked eye, containing a large number of cells. Conclusions: The developed protocol can be useful for application of somatic hybridization to improve date palm cultivars.https://www.ijbiotech.com/article_10558_9029be790b5f881ef6b973a5a94d5f94.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304313120150301Identification of Crocus sativus and its Adulterants from Chinese Markets by using DNA Barcoding Technique3642981110.15171/ijb.1034ENWeijuan HuangDepartment of Biology, College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, PR ChinaFeifei LiDepartment of Biology, College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, PR ChinaYujing LiuDepartment of Biology, College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, PR ChinaChunlin Long1-Department of Biology, College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, PR China
2-Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, PR ChinaJournal Article20141007Background: Saffron (Crocus sativus L.) is a common but very expensive herbal medicine. As an important traditional medicine, it has an outstanding effect in treating irregular and painful menstruation. Recently, the over-demand tendency of saffron results in an unusual phenomenon in the medicinal markets. Adulterants and saffron-like substitutes are intentionally mixed into medicinal markets and pharmacies or online stores, affecting drug safety and food quality. Objectives: Our study aimed to identify saffron from its adulterants via DNA barcoding. Materials and Methods: Samples (13 saffron + 4 others containing Carthamus tinctorius or Chrysanthemum x morifolium) obtained from 12 different provinces of China. Through DNA barcoding, samples were compared using three candidate markers, trnH-psbA, rbcL-a and ITS2. Results: trnH-psbA and rbcL-a were capable of distinguishing different accessions. ITS2 could identify samples even at intra-specific level. According to these three barcodes, four samples were identified saffron-like substitutes. Conclusions: The adulterant rate in Chinese markets reaches as high as 33.33% that may cause health risks and further may reduce saffron efficacy once is being used as herbal remedy. In order to make a distinction between C. sativus with other genera as adulterants, DNA barcoding is suggested.https://www.ijbiotech.com/article_9811_5b7b37efebae69acf10cbca26bd8119e.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304313120150301The Association of Bovine Osteopontin (OPN) Gene with Milk Production Traits in Iranian Holstein Bulls4348808610.15171/ijb.1092ENAbdolreza SalehiDepartment of Animal and Poultry Science, College of Aburaihan, University of Tehran, Tehran, IranKhadijeh NasiriDepartment of Animal and Poultry Science, College of Aburaihan, University of Tehran, Tehran, IranMahdi AminafsharDepartment of Animal Science, Faculty of Agriculture and Natural Resourse, Science and Research Branch, Islamic Azad University, Tehran, IranRohoallah SobhaniDepartment of Animal and Poultry Science, College of Aburaihan, University of Tehran, Tehran, IranMohammad Bagher SayaadnejadAnimal Breeding Centre of Iran, Karaj, IranJournal Article20141202Background: The Osteopontin (OPN) is a highly phosphorylated glycoprotein in numbers of bovine tissues and milk. OPN has been reported to be associated with milk production in cattle. Objective: The genotype and allelic frequencies for OPN and its association with milk production will be evaluated in Iranian Holstein Bulls. Materials and Methods: Bulls DNA (100) was isolated. Oligo was used for primer design. Polymerase Chain Reaction was implemented to amplify a 826 bp fragment and the amplicon was digested by BsrI. Restricted Maximum likelihood (REML) method based on average information algorithm using ASRMEL programs (version 3.1) was employed to estimate the genetic parameters and variance of components. The association of OPN genotypes with milk production traits were analysed by the least square method as applied in the general linear model (GLM) procedure of SAS. Allele substitution effects were performed by regression analyses. Results: Allele frequencies of T and C were 0.59±0.03 and 0.41±0.03, respectively. Genotype frequencies of TT, CT and CC were 34.69, 48.62, and 16.69, respectively. The chi-square test showed the deviation from Hardy-Weinberg equilibrium. Estimated heritability for milk yield, fat yield and its percent, protein yield and its percent were 0.28±0.0061, 0.21±0.0064, 0.22±0.0086, 0.32±0.0065 and 0.34±0.0096 respectively. Allelic substitution effects and differences between genotypes were not significant for milk production traits. Conclusions: This study suggested that the C allele frequency of OPN was noticeable in Iranian proven bull Holstein population, but was not associated with milk production traits. However, before being practical for the breeding improvement of Iranian Holsteins a larger sample size is required.https://www.ijbiotech.com/article_8086_81e690fbdad35a4c2cd9e92a61dbc82d.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304313120150301Association of Two Polymorphic Codons in P53 and ABCC1 Promoter with Prostate Cancer49541055910.15171/ijb.1096ENFarinaz BehfarjamDepartment of Biology, Faculty of Science, University of Kurdistan, Sanandaj, IranJalal RostamzadehDepartment of Animal Sciences, Faculty of Agriculture, University of Kurdistan, Sanandaj, IranMohammad Ali ZareiDepartment of Biology, Faculty of Science, University of Kurdistan, Sanandaj, IranBahram NikkhooFaculty of Medicine, Kurdistan University of Medical Science, Sanandaj, IranJournal Article20141206Background: In prostate cancer, mutated p53 alleles typically contain missense single-base substitution in codon 72 that resides within exons 5-8. Stable p53 proteins in tumor cell nuclei have been associated with malignancy. A role of p53 is the regulation of drug transporters like ABCC1 (MRP1) by an effect on promoter region. Objectives: The objective of this study was to identify association of mutations of p53 at codon 72 and 282 and promoter region of ABCC1 with increased risks of prostate cancer. Materials and Methods: Formalin fixed, paraffin-embedded malignant tissues of 45 patients and 45 control samples were evaluated. PCR-RFLP using BstUI for codon 72 and HpaII restriction enzyme for codon 282 p53 gene, and G-1666A promoter region of ABCC1 gene was performed. To assess the frequency of these mutations and to detect new mutations in cancerous samples, PCR-SSCP analysis was performed. Results: The frequencies of CC, GC and GG genotypes of codon 72 of p53 were 33.33%, 46.67% and 20.00% in patients with cancer and 15.56%, 48.89% and 35.55% in controls, respectively. The relative allele frequencies of ABCC1 promoter polymorphism were 60.00% A and 40.00% G in patients as opposed to 37.78% for A and 62.22% for G in controls. Genotypic frequencies of p53 codon 72 and G1666A of ABCC1 in patients vs. Controls were statistically significant(p<0.05). The study of these samples with PCR-SSCP displayed some new banding patterns. Conclusions: The present findings suggest that CC homozygosity in codon 72 of p53 gene and AA genotype in G-1666A of ABCC1 gene may play a role in combination in prostate cancer and increased susceptibility for this malignancy in the Iranian Kurdish population.https://www.ijbiotech.com/article_10559_5f3f55903d0474bd001b4039b72266f1.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304313120150301Evaluation of Cell Penetrating Peptide Delivery System on HPV16E7 Expression in Three Types of Cell Line5562980910.15171/ijb.1115ENTayebeh SalehDepartment of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, IranAzam BolhassaniDepartment of Hepatitis and AIDs, Pasteur Institute of Iran, Tehran, IranSeyed Abbas ShojaosadatiBiotechnology Group, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran0000-0002-8561-2414Saman HosseinkhaniDepartment of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran0000-0002-0345-7909Journal Article20141227Background: The poor permeability of the plasma and nuclear membranes to DNA plasmids are two major barriers for the development of these therapeutic molecules. Therefore, success in gene therapy approaches depends on the development of efficient and safe non-viral delivery systems. Objectives: The aim of this study was to investigate the in vitro delivery of plasmid DNA encoding HPV16 E7 gene using cell penetrating peptide delivery system to achieve the best conditions for cell transfection and protein expression. For this purpose, we have used a cationic peptide delivery system, MPG which forms stable non-covalent complexes with nucleic acids for delivery of pEGFP-E7 as a model antigen in vitro. Materials and Methods: DNA construct encoding HPV16 E7 (pEGFP-E7) was prepared in large scale with high purity. MPG peptide/ DNA complexes were prepared at different N/P (nitrogen/phosphate) ratios and physicochemical characterization and stability of nanoparticles were investigated. In vitro peptide-mediated E7-GFP DNA transfection, and its expression was evaluated in three cell types. To quantify the transfection efficiency of this delivery system, transfected cells were harvested and assessed for GFP-positive cells by flow cytometry. Furthermore, E7-GFP expression was confirmed by western blot analysis. Results: The cellular uptake of MPG based nanoparticles was shown to be comparable with standard reagent PEI. The COS-7 cells transfected by MPG-based nanoparticles at an N/P ratio of 15:1 showed the highest transfection efficiency and gene expression. Conclusions: The results indicated that the efficient gene expression depends on both cell type and N/P ratio applied, in vitro. The efficient protein expression detected by western blotting and flow cytometry supports the potential of MPG-based nanoparticles as a potent gene delivery system.https://www.ijbiotech.com/article_9809_715cf7e13cd8b234e2e8fa81d08b219b.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-304313120150301Detection of VIM- and IMP-type Metallo-Beta-Lactamase Genes in Acinetobacter baumannii Isolates from Patients in Two Hospitals in Tehran6367792710.15171/ijb1088ENSaba DavoodiDepartment of Biology, Science and Research Branch, Islamic Azad University, Tehran, IranMohammad Ali BoroumandDepartment of Molecular Pathology, Tehran Heart Center, Tehran University of Medical Science, Tehran, IranSaeed SepehrisereshtDepartment of Molecular Pathology, Tehran Heart Center, Tehran University of Medical Science, Tehran, IranLeila PourgholiDepartment of Molecular Pathology, Tehran Heart Center, Tehran University of Medical Science, Tehran, IranJournal Article20141127Background: Acinetobacter baumannii, is an opportunistic pathogen and is responsible for numerous nosocomial infections. In recent years, this microorganism has been resistant to a wide range of antibiotics. One of the most important mechanisms of resistance in this microorganism is production of metallo-beta-lactamases (MBLs). Objectives: The aim of this study was to detect VIM- and IMP-type metallo-beta-lactamase genes in Acinetobacter baumanniiisolates from patients in two Hospitals in Tehran. Materials and Methods: 104 isolates were tested using the PCR method for the identification of VIM- and IMP-type genes. Results: vim1, vim2, imp1and imp2 genes were detected in 6.7%, 41.7%, 50% and 1.7% of the isolates from Tehran Heart Center, and in 29.5%, 38.6%, 4.5% and 4.5% of the isolates from Shahid Mutahhari Hospital respectively. Discussion: Our analysis revealed that the majority of the isolates had at least one of these genes, indicating that MBLs production is an important resistance mechanism in Acinetobacter baumannii.https://www.ijbiotech.com/article_7927_3f72bb55e5d5b09823212bb44089b157.pdf