Ferdowsi University of Mashhad
Journal of Cell and Molecular Research
2008-9147
2717-3364
8
2
2016
12
01
Association of Codon 72 of P53 Gene Polymorphism with Chronic Hepatitis C Virus Infection: A Case Control Study
46
51
EN
Sina
Gerayli
Western University
sina.gerayli@gmail.com
Alireza
Pasdar
University of Aberd
a.pasdar@abdn.ac.uk
Sina
Rostami
University of Bergen
sin.fum@gmail.com
Samaneh
Sepahi
Mashhad University of Medical Sciences
samanehsepahi@yahoo.com
Seyed Mousalreza
Hoseini
Mashhad University of Medical Sciences
hoseinimr@mums.ac.ir
Reza
Jahanian
Mashhad University of Medical Sciences
Aida
Gholoobi
Mashhad University of Medical Sciences
Zahra
Meshkat
Mashhad University of Medical Sciences
meshkatz@mums.ac.ir
Mitra
Ahadi
Mashhad University of Medical Sciences
ahadim@mums.ac.ir
10.22067/jcmr.v8i2.56120
Single nucleotide polymorphism in codon 72 of p53 gene (Arg/Pro) changes p53 protein structure and affects its activities. Hepatitis C virus (HCV) is believed to induce hepatocellular carcinoma and P53 polymorphisms have been associated with human cancers. The aim of this study was to evaluate genetic variants of codon 72 of p53 gene polymorphism in HCV patients and its relationship with HCV infection.
The study was conducted on 67 HCV patients, who were referred to medical centers of Mashhad city, Iran, and 73 healthy people from the same region. Genotyping of codon 72 of p53 gene was performed by PCR-RFLP method.
The distributions of different alleles of p53 polymorphisms did not differ significantly between groups. The respective proportions of Proline homozygotes, heterozygotes, and Arginine homozygotes were 37.31%, 35.82%, 26.86% in patients and 39.72%, 27.39%, and 32.87% in the control group respectively. However, we found no significant differenece for the allelic or genotype distribution between cases and controls.
Our results indicated no strong evidence of association of the p53 polymorphism with HCV infection; however, further investigation is needed in different ethnic groups to elucidate the role of this polymorphism in HCV infection.
Polymorphism,P53 gene,HCV,Genetic epidemiology,Iran
https://jcmr.um.ac.ir/article_27680.html
https://jcmr.um.ac.ir/article_27680_a2195e2e26876b9425bb72a2a64b9af3.pdf
Ferdowsi University of Mashhad
Journal of Cell and Molecular Research
2008-9147
2717-3364
8
2
2016
12
01
Geometric Mean of 5S rRNA and MiR-16 As A Suitable Normalizer In Esophageal Cancer
52
57
EN
Samaneh
Khazaei
Isfahan University
khazaei.samaneh@yahoo.com
Sedigheh
Gharbi
shahid Bahonar university
magharbi@gmail.com
Seyed Javad
Mowla
0000-0002-3300-6332
Tarbiat modares university
sjmowla@modares.ac.ir
10.22067/jcmr.v8i2.56251
Esophageal squamous cell carcinoma (ESCC) is a deadly cancer with poor prognosis. In this regard, early diagnosis is of vital importance to cure the tumor in its early stages. Novel cancer diagnostic and therapeutic approaches have been recently introduced based on microRNAs (miRNAs). Also, accurate normalization using appropriate reference genes is a critical step in miRNA expression studies. In this study, we aimed to identify appropriate reference genes for miRNA quantification in serum samples of ESCC. In this case and control experimental study, two statistical algorithms including GeNorm and NormFinder were used to evaluate the suitability of miR-16 and 5S rRNA and their geometric mean as reference genes. Then, relative expression of miR-451 and miR-24 were evaluated while different normalizer including miR-16, 5S rRNA and their geometric mean were applied. Both GeNorm and NormFinder analyses showed that geometric mean of miR-16 and 5S rRNA is the most stable reference gene in these samples. Also, our data showed that choosing an inappropriate normalizer could change the relative expression of target genes of miR-451 and miR-24 in ESCC samples which emphasize on the importance of selecting a reliable internal control in expression analyses. We demonstrated that geometric mean of two reference genes could increase the reliability of normalizers and also by using geometric mean as reference gene, relative expression of different target is closer to reality.
Esophageal cancer,microRNA,qRT-PCR,Reference genes
https://jcmr.um.ac.ir/article_27711.html
https://jcmr.um.ac.ir/article_27711_ce1fdadde9af8bdc53017ab03314e7b1.pdf
Ferdowsi University of Mashhad
Journal of Cell and Molecular Research
2008-9147
2717-3364
8
2
2016
12
01
Protective Effect of Diosgenin against H2O2-Induced Oxidative Stress on H9C2 Cells
58
64
EN
Samaneh
Jamshidi
Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Iran
samaneh_1834@yahoo.com
Mehrdad
Lahouti
Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Iran
mlahouti@um.ac.ir
Mohammad Taher
Boroushaki
Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Science, Mashhad, Iran
boroushakimt@mums.ac.ir
Ali
Ganjeali
0000000209568650
Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Iran
ganjeali@um.ac.ir
Ahmad
Ghorbani
Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Science, Mashhad, Iran
Mehdi
Bihamta toosi
Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Science, Mashhad, Iran
meh_ts@yahoo.com
10.22067/jcmr.v0i0.36148
Diosgenin is an important compound in pharmaceutical industry. It has various effects such as hypocholesterolemic action or antioxidant activity in HIV infected patients. Biological oxidation pathways are involved in causing or aggravating heart disease. This study investigated the potential protective effect of diosgenin on cell viability and antioxidant defenses of cultured H9C2 cells submitted to oxidative stress induced by H2O2. Viability of cells exposed to H2O2 was detected by MTT assay. The generation of ROS and hydrogen peroxide release after H2O2 were detected using the fluorescent probe H2DCF-DA. The lipid peroxidation product i.e. MDA formation was estimated by assessing the levels of thio-barbituric acid reactive substances (TBARS) using spectrophotometry. SOD activity was assayed with NWLSS (TM) Superoxide Dismutase (SOD) activity assay kit. Pretreatment of cells with 3-25 µM of diosgenin for 24 h before applying H2O2 completely prevented cell damage and significantly enhanced viability of H9C2 cells. Increased ROS induced by H2O2 was dose dependently prevented when cells were pretreated for 24 h with diosgenin. The level of the lipid peroxidation was significantly higher in H9C2 cells exposed to H2O2 as compared to the control and cells pretreated with diosgenin. SOD activity in cells treated with diosgenin significantly decreased compared with cells exposed to H2O2. These results show that treatment of H9C2 cells with diosgenin (3-25 µM) confers a significant protection against oxidative stress.
Diosgenin,H9C2 cells,oxidative stress,MDA,Cell viability
https://jcmr.um.ac.ir/article_27742.html
https://jcmr.um.ac.ir/article_27742_5411cc219fc888c6ed59167ccc8116c2.pdf
Ferdowsi University of Mashhad
Journal of Cell and Molecular Research
2008-9147
2717-3364
8
2
2016
12
01
Construction of the Recombinant Lentiviral Vector Containing Human GH1 Gene and Its Expression in HEK293T Cells
71
77
EN
Zahra
Roudbari
Ferdowsi University of MashhadUniversity of Mashhad
rodbari.zahra@gmail.com
Mohammadreza
Nassiri
Ferdowsi University of Mashhad
nassiryr@um.ac.ir
Mojtaba
Tahmoorespur
Ferdowsi University of Mashhad
m_tahmoorespur@yahoo.com
Aliakbar
Haddad-Mashadrizeh
Ferdowsi University of Mashhad
aliakbar.haddad@gmail.com
Ali
Javadmanesh
0000-0001-6016-5905
Ferdowsi University of Mashhad
javadmanesh@um.ac.ir
10.22067/jcmr.v8i2.22857
Human growth hormone (hGH) is a protein with multiple roles in a range of biological functions such as protein, carbohydrates and lipid metabolisms as well as immunity, tissue development and overall growth. One of the major class of biopharmaceuticals in mammalian cells is the production of recombinant pharmaceutical proteins. In this study, we constructed a lentiviral vector carrying coding region of human GH1 (hGH) gene in order to production of recombinant hGH in mammalian cell line. hGH gene was amplified from a plasmid containing full-length hGH coding sequence and then cloned into the lentiviral vector pCDH-GFP. The HEK293T cells were transduced by the lentivirus particles as a targeted cell. hGH expression status in the recombinant cells were confirmed by RT-PCR. Additionally, western blotting analysis results showed that the recombinant cells maintained a stable hGH expression during five weeks of continuous culture. In conclusion, results of current study suggested that constructed lentiviral vector can potentially be used for a stable production of recombinant hGH protein in HEK293T cells. This methodology could be served as a foundation for further research and may open new insights toward therapeutic protein manufacturing.
hGH,Recombinant lentivirus,Production protein,HEK 293 cells
https://jcmr.um.ac.ir/article_27765.html
https://jcmr.um.ac.ir/article_27765_2d0acd9ae2a704eb8fab22493bb197d6.pdf
Ferdowsi University of Mashhad
Journal of Cell and Molecular Research
2008-9147
2717-3364
8
2
2016
12
01
Unravelling Over-Represented Amino Acids in Protein Structure of Allergen Proteins; A Large-Scale Study
65
70
EN
Nassim
Rahmani
Shiraz University
rahmani_nassim@yahoo.com
Esmaeil
Ebrahimie
Shiraz University
esmaeil.ebrahimie@adelaide.edu.au
Ali
Niazi
Shiraz University
niazi@shirazu.ac.ir
Najaf
Allahyari Fard
National Institute of Genetic Engineering and Biotechnology (NIGEB)
allahyar@nigeb.ac.ir
Bijan
Bambai
National Institute of Genetic Engineering and Biotechnology (NIGEB)
bambai@nigeb.ac.ir
Zarrin
Minuchehr
National Institute of Genetic Engineering and Biotechnology (NIGEB)
minuchehr@nigeb.ac.ir
Mansour
Ebrahimi
University of Qom
mansour@future.edu
10.22067/jcmr.v0i0.60190
Allergens are proteins or glycoproteins which make widespread disorders that can lead to a systemic anaphylactic shock and even death within a short period of time. Understanding the protein features that are involved in allergenicity is important in developing future treatments as well as engineering proteins in genetic transformation projects. A big dataset of 1439 protein features from 761 plant allergens and 7815 non-allergen proteins was constructed. Thereafter, 10 different attribute weighting algorithms were utilized to find the key characteristics differentiating allergens and non-allergen proteins. The frequency of Leu, Arg and Gln selected by different attribute weighting algorithms with more than 50% confidence, including attribute weighting by Weight_Info Gain, Weight Chi Squared, Weight_Gini Index and Weight_Relief. High amount of Gln and low percentage of Leu and Arg discriminate plant allergens from non-allergens
Plant allergens,Attribute weighting algorithms,Amino acid
https://jcmr.um.ac.ir/article_27773.html
https://jcmr.um.ac.ir/article_27773_d730e67f27bf04957720ffbfbcc2aa6f.pdf
Ferdowsi University of Mashhad
Journal of Cell and Molecular Research
2008-9147
2717-3364
8
2
2016
12
01
Transient Expression of Coat Protein of Foot and Mouth Disease Virus (FMDV) in Alfalfa (Medicago sativa) by Agroinfiltration
83
89
EN
Maziar
Habibi-Pirkoohi
Ferdowsi University of Mashhad
maziar.habibi.p@gmail.com
Saeid
Malekzadeh-Shafaroudi
Department of Biotechnology and plant breeding, Ferdowsi University of Mashhad, Mashhad, Iran
malekzadeh-s@um.ac.ir
Hasan
Marashi
Department of Biotechnology and plant breeding, Ferdowsi University of Mashhad, Mashhad, Iran
marashi@ferdowsi.um.ac.ir
Saeid
Zibaee
Razi Vaccine and Serum Research Institute, Ministry of Agriculture, Mashhad, Iran
s.zibaee@mrazi.ac.ir
Afsaneh
Mohkami
Horticulture research Institute, Shahid Bahonar University, Kerman, Iran
afi_mohkami@yahoo.com
Saba
NejatizaXeh
Department of Biotechnology and plant breeding, Ferdowsi University of Mashhad, Mashhad, Iran
s.siklamen@yahoo.com
10.22067/jcmr.v8i2.44884
An Agrobacterium-mediated transient gene expression assay was carried out in alfalfa (Medicago sativa) leaves for expression of a chimeric gene encoding a part of capsid protein of Foot and Mouth Disease virus called VP1. The plant leaves were transformed via agroinfiltration procedure. The presence of the foreign gene and its expression in transformed plants were evaluated by polymerase chain reaction (PCR), real time PCR, protein Dot blot and ELISA. Moreover, gene expression in the transformed leaves was quantified by ELISA method. The results obtained in this investigation indicated high level of gene expression in alfalfa leaves, showing that transient gene expression can be applied as an effective and time-saving procedure for the production of recombinant proteins. The procedures for transformation, detection of recombinant protein and its application for molecular experiments are described in the study.
Agroinfiltration,FMDV,Recombinant vaccine,Alfalfa,VP1
https://jcmr.um.ac.ir/article_27794.html
https://jcmr.um.ac.ir/article_27794_24c75230f71999bd6ae27242c143be33.pdf
Ferdowsi University of Mashhad
Journal of Cell and Molecular Research
2008-9147
2717-3364
8
2
2016
12
01
Purification and Characterization of an Extracellular Phosphatase Enzyme From Bacillus spp.
90
97
EN
Maryam
Parhamfar
Shahid Bahonar University of Kerman
mary_parhamfar@yahoo.com
Arastoo
Badoei-Dalfard
Shahid Bahonar University of Kerman
badoe@uk.ac.ir
Milad
Parhamfar
Duissburg-Essen University
milad.parhamfar@stud.uni-due.de
Shohreh
Fahimi Rad
Campus of Agriculture and Natural Resources, University of Tehran
sh_fahimirad@yahoo.com
10.22067/jcmr.v8i2.58676
Phosphorus is one of the most important nutrients for plant growth and development. Chemical Pi fertilizer is used to provide the phosphorus for the plants, but it is mostly fixed in the soil into insoluble form and become unavailable to the plants. Phosphate-solubilizing bacteria have lots of application in agriculture as biological fertilizer. Consumption of biofertilizers instead of chemical fertilizers can lead to environmental pollution reduction and crop production enhancement using sustainable farming. In this study, a phosphatase-producing bacterium was isolated from agricultural soil in Kerman. Screening of phosphate solubilizing bacteria was performed on the PVK medium, based on clear area diameter. The best bacterium (AG41) was identified based on 16s rDNA gene. The optimum condition for production of phosphatase was also determined and it was purified and characterized. Sequence alignment and phylogenetic tree results show that AG41 is closely related to Bacillus subtilis, with 98% homology. Phosphatase activity was determined by end point method. The best carbon, nitrogen and phosphate sources for enzyme production were 1.0% glucose, 0.5% ammonium sulfate and (0.25%) sodium phytate +(0.25%) tricalcium phosphate, respectively. Bacterial phosphatase was partially purified using ammonium sulfate fractionation followed by dialysis. Results showed that the optimum temperature for the purified enzyme activity was 40oC and it was stable at temperatures below 60°C. This enzyme was stable between pH 3.0-7.0, and the optimal pH activity was found to 5.0. These results indicated that this strain can be a notable candidate for using as biofertilizers.
Screening,Biofertilizer,Phosphate-solubilizing bacteria,Phosphatase
https://jcmr.um.ac.ir/article_27806.html
https://jcmr.um.ac.ir/article_27806_7efa22ceeeaf08b84da619e93937b1ee.pdf