Hepcidin25 is a small cysteine-rich peptide hormone known as a new class of antimicrobial peptide... more Hepcidin25 is a small cysteine-rich peptide hormone known as a new class of antimicrobial peptides. The purpose of the present study was to express, purify and investigate the antibacterial properties of recombinant human hepcidin25 protein production in Escherichia coli. Human hepcidin25 gene was optimized and fused to a small ubiquitin-related modifier (SUMO) gene for higher expression. Then SUMO-hepcidin25 was cloned into the pET-32a (+) vector and expressed in E.coli Origami. The fusion protein with a molecular weight of approximately 35 kDa was analyzed on SDS-PAGE gel. The highest expression was observed after 6 hours induction and the fusion protein consisted approximately 47 % of the total cellular protein. The purified SUMO-hepcidin25 purity was determined to be higher than 95 %, with a final yield of 3.9 mg L-1 of media. The recombinant hepcidin25 showed antibacterial activity against both gram negative (Klebsiella pneumonia) and gram positive)Staphylococcus aureus and Bacillus cereus(bacteria with minimum inhibitory concentrations (MICs) of 150 µg mL −1 , 18.7 µg/mL −1 and 37.5 µg/mL −1 , respectively. These results indicated that thioredoxin and SUMO dual fusion system is an efficient production system for synthesis functional human hepcidin25.
Background: In recent decades RNAi has become a novel and effective tool to silence gene expressi... more Background: In recent decades RNAi has become a novel and effective tool to silence gene expression. The expression of shRNAs against target genes has now become a standard and powerful strategy for antiviral therapeutic approaches. Infectious bovine rhinotracheitis (IBR) is a domestic and wild ruminant viral disease that caused by bovine herpesvirus-1. Occasionally, its entry into the herd may cause vigorous economic damage due to losing weight, reduced output and limitation on livestock international trade. Vaccines against the disease are not fully effective and the need for an effective therapeutic approach is highly felt. In the present study an RNAi-based gene therapy was designed and implemented. Due to the essential role of UL25 gene for BHV-1 replication. Method: the sequence of UL25 gene was purposed for designing shRNA molecules. The UL25 gene sequence was extracted from the NCBI database and suitable shRNA molecules were purposed by using online soft wares. Three recombinant lentiviral vectors expressing short hairpin RNAs (shRNAs) versus the UL25 gene of IBRV were constructed and co-transfected into HEK 293T cells. The effectiveness of designed shRNA molecules was assayed by calculating TCID50 titers and observation of BHV-1 cytopathic effects. Results: The nal calculations showed that all shRNAs had antiviral effects. In the TCID50 test, the shRNAs 1, 2 and 3 su ciently decreased BHV1 output in comparison with the control groups (almost 98.22%, 99.63% and 99.54%, respectively than cells with IBRV inoculation and nearly 90.84%, 97.9% and 96.84%, respectively than cells with ORFV and scrambled vector inoculation). ShRNA-2 achieved maximal inhibition of viral replication. Conclusion: The results indicated that shRNAs targeting the UL25 gene showed considerable antiviral attributes and reduced IBRV multiplication in MDBK cells. In conclusion; RNA interference can be used as a gene therapy instrument against BHV-1 virus.
International Journal of Peptide Research and Therapeutics, Aug 16, 2022
Anti-microbial peptides (AMPs) are important factors of the innate immune system which perform a ... more Anti-microbial peptides (AMPs) are important factors of the innate immune system which perform a comprehensive spectrum of biological activities. The present study aims to examine the expression, puri cation, and cytotoxic effects of recombinant PI-laterosporulin10 (LS10) a defensing-like peptide. PI is a transmembrane peptide that helps to increase penetration and speci city of peptide targeting. PI-LS10 sequence was optimized and fused to a Xa protease site to achieve higher levels of expression. Then Xa-PI-LS10 was cloned into the pET-32a (+) vector and expressed in E. coli BL21 (DE3). The fusion protein with a molecular weight of about 25.3 kDa was analyzed using the SDS-PAGE. The maximum expression was observed 8 hours post-induction. After enzymatic cleavage, the purity of PI-LS10 was >300 µg/ ml as the nal yield. The recombinant PI-LS10 showed anti-cancer characteristics against breast cancer cell lines. MCF7 (non-triple negative) and MDA-MB-231 (triple negative) cells were treated with different concentrations of the PI-LS10 in order to evaluate their cytotoxic activity and e ciency in targeting triple negative cells. Our results suggest that PI-LS10 had a signi cant cytotoxic effect against breast cancer cells in comparison to the normal cell line. Triple negative breast cancer cell (MDA-MB-231) proliferation inhibited higher in 72 hours after treatment compare MCF7 cells. In conclusion, the PI-LS10 peptide was produced successfully and its favorable anti-cancer activity against a breast cancer cell line was con rmed.
The production of natural antibiotic peptides has emerged as an important mechanism of innate imm... more The production of natural antibiotic peptides has emerged as an important mechanism of innate immunity in plants and animals. Defensins are diverse members of a large family of antimicrobial peptides, contributing to the antimicrobial action of granulocytes, mucosal host defence in the small intestine and epithelial host defence in the skin and elsewhere. This review, inspired by a spate of recent studies of defensins in human diseases and animal models, focuses on the biological function of defensins.
Background and Aims: Multicellular organisms immune system has less than 100 amino acid peptide. ... more Background and Aims: Multicellular organisms immune system has less than 100 amino acid peptide. Defensins are one of the largest family of these antimicrobial peptides and because it is applicable as much to the benefit of new generations of antibiotics. The purpose of this study was to evaluate gene expression of beta defensin 2 this is antimicrobial peptides. Methods: In this study the gene sequences BNBD2 used considering codons in E coli. and in expression vector pET-32a (+) was sub cloned. E.coli strain BL21 by the recombinant vector transfection and protein expression under IPTG induction and SDS-PAGE electrophoresis and Western blot was evaluated. Results: Recombinant proteins were expressed with IPTG. Molecular weight on gel electrophoresis (KD25) with molecular weight equal to that predicted by theoretical calculations. Techniques western blotting confirmed the presence of this protein. Conclusions: This study confirmed the sub-cloning and expression BNBD2. The high activity of antimicrobial peptides in vitro in the near future can be effective in treating bacterial infections in vivo.
Journal of the Hellenic Veterinary Medical Society, Jan 28, 2022
In silico design of short hairpin RNA (shRNA)Molecules for DNA pol gene of Contagious Ecthyma vir... more In silico design of short hairpin RNA (shRNA)Molecules for DNA pol gene of Contagious Ecthyma virus (ORFV)
Background: Border disease is believed to be one of the most important diseases in the animal hus... more Background: Border disease is believed to be one of the most important diseases in the animal husbandry industry, which has not yet been eradicated in Iran. The development of approaches based on the application of interfering RNA (RNAi) for antiviral therapy has attracted a great deal of attention over the recent years. The present research was conducted to design, construct, and apply shRNA against the NS3 gene of BDV to evaluate the prevention of BDV proliferation in the cell culture system. For this purpose, the suitable oligonucleotide sequence of NS3 gene coding was selected utilizing BDV-X818 strain. Afterwards, using shRNA design software, shRNA molecules were designed and synthesized. These shRNAs were cloned into the desired vectors and were finally transfected in HEK293T cells employing the third generation of lentiviral packaging system. Subsequently, these shRNA expressing lentiviruses were transduced to the MDBK cell line to challenge to border virus. In order to evaluate the efficacy of shRNAs, the viral infectious titer and RNA copy number were calculated with TCID50 and Real-time RT-PCR tests, respectively. Results: The results revealed that shRNAs 1, 2, and 3 decreased viral RNA by more than 90% compared to the control groups. BDV titer noticeably decreased after the challenge with shRNAs 1, 2, and 3 from ∼88% up to 99% in comparison with the control groups. Conclusions: Overall, it could be concluded that RNAi may be considered as a strong treatment proposal against viruses, such as BDV.
Background and purpose: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscul... more Background and purpose: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder. Carrier frequency studies of SMA have been reported for various populations. Although no large-scale population-based studies of SMA have been performed in Iran, previous estimates have indicated that the incidence of autosomal recessive disorder partly because of the high prevalence of consanguineous marriage is much higher in the Iranian population than in other populations. Methods: In this study, we used a reliable and highly sensitive quantitative real-time PCR assay with SYBR green I dye to detect the copy number of the SMN1 gene to determine the carrier frequency of SMA in 200 healthy unrelated, non-consanguineous couples from different part of Iran. Results: To validate the method in our samples, we determined the relative quantification (RQ) of patients with homozygous deletion (0.00) and hemyzygous carriers (0.29-0.55). The RQ in 10 of 200 normal individuals were within the carrier range of 0.31-0.57, estimating a carrier frequency of 5% in the Iranian population. Conclusions: Our data show that the SMA carrier frequency in Iran is higher than in the European population and that further programs of population carrier detection and prenatal testing should be implemented.
Jentashapir Journal of Cellular and Molecular Biology
Border disease virus (BDV) is a serious pathogen of sheep and goats worldwide. The virus causes c... more Border disease virus (BDV) is a serious pathogen of sheep and goats worldwide. The virus causes considerable economic losses in animal husbandry. Therefore, developing effective and genetic-based therapies to control viral infections, such as BDV, has been remarkable over recent years. A common way to evaluate such therapeutic strategies is by cloning the desired fragment into appropriate vectors to develop cell lines expressing it. Due to the required duties of the NS3 gene for BDV proliferation, this gene's HELICc and nucleotide binding site domain was synthesized and cloned in a lentiviral vector upstream of the GFP gene. The cloning accuracy was verified by digestion with restriction enzymes and sequencing. Thus, the BDV-NS3 HELICc and nucleotide binding site carrying plasmid was prepared successfully and will be applied in a second or third lentiviral packaging system for stable expression of the desired gene.
Journal of the Hellenic Veterinary Medical Society
Contagious ecthyma is an infectious skin disease of ruminants caused by the ORF virus (ORFV) that... more Contagious ecthyma is an infectious skin disease of ruminants caused by the ORF virus (ORFV) that is a member of genus Parapoxvirus of the Poxviridae family. In addition to the significant effects on lambs and human, ORFVs have been recently shown to infect other hosts. The disease causes significant economic damages to the sheep industry, so attempts to eliminate it must be taken into.RNA interference (RNAi) is an evolutionarily conserved mechanism in which the expression of homologous target genesis is suppressed by means of double-stranded RNA molecules. Since RNAi can be considered as a therapeutic method for viral gene silencing, we tend to make the most of this capability. The present study aims to design potential shRNAs to knockdown the DNA-polymerase gene coded by ORF025.A significant number of computational methods such as clustal omega website to target alignment, BLAST-NCBI to similarity search, CLC software to secondary structure prediction, BLOCK-iTRNAi Designer and WI...
Anti-microbial peptides (AMPs) are important factors of the innate immune system which perform a ... more Anti-microbial peptides (AMPs) are important factors of the innate immune system which perform a comprehensive spectrum of biological activities. The present study aims to examine the expression, purification, and cytotoxic effects of recombinant PI-laterosporulin10 (LS10) a defensing-like peptide. PI is a transmembrane peptide that helps to increase penetration and specificity of peptide targeting. PI-LS10 sequence was optimized and fused to a Xa protease site to achieve higher levels of expression. Then Xa-PI-LS10 was cloned into the pET-32a (+) vector and expressed in E. coli BL21 (DE3). The fusion protein with a molecular weight of about 25.3 kDa was analyzed using the SDS-PAGE. The maximum expression was observed 8 hours post-induction. After enzymatic cleavage, the purity of PI-LS10 was >300 µg/ ml as the final yield. The recombinant PI-LS10 showed anti-cancer characteristics against breast cancer cell lines. MCF7 (non-triple negative) and MDA-MB-231 (triple negative) cells...
The author regrets that, in the article titled ''Functional analysis of recombinant codon-optimiz... more The author regrets that, in the article titled ''Functional analysis of recombinant codon-optimized bovine neutrophil b-defensin", he missed to include the details of the reference in the legend of Fig. 5. The details of the missed reference is as follows: ''Karimi N, Saffar B, Ghaedi K, Mobini M. ''Optimization of expression, purification and handling anti-bacteria feature protein of bovine neutrophil beta defensin 2 in E. coli."
Background: Colorectal cancer (CRC) is one of the most common cancers in the world. High activity... more Background: Colorectal cancer (CRC) is one of the most common cancers in the world. High activity of CyclinD1 gene has been seen in the progress of this cancer. A common polymorphism (G870A) in exon 4 of CyclinD1 produces a variant transcript with longer half-life and may cause uncontrollable cellular growth thus contributing to cancer development. This study was performed to evaluate the frequency of CCND1 G870A polymorphism between CRC cases and controls. Methods: DNA samples from peripheral blood leukocytes of 50 patients with sporadic colorectal cancer and 50 healthy subjects were extracted. CCND1 G870A polymorphism was genotyped by polymerase chain reaction -restriction fragment length polymorphism (PCR-RFLP) in healthy subjects and patients. Statistical analysis was performed by chi-squared test via SPSS software. Findings: Relationship between CCND1 G870A polymorphism in allele frequencies between cases and controls were not observed (P = 0.204). While the frequency of AA gen...
Introduction:The amylase enzyme hydrolyze starch and break glycosidic internal bonds of this poly... more Introduction:The amylase enzyme hydrolyze starch and break glycosidic internal bonds of this polysaccharides. Amylases are important industrial enzymes for a wide range of applications including starch conversion to sugar syrup, dextrin production, and use in the pharmaceutical and medical industries. Materials and methods: In this study, thermophilic bacterium was isolated and purified from the Qynarcheh hot spring in Ardabil. Desired strain suspension in nutrient broth medium was placed in front of UV rays with a wavelength of 254 nm at a distance of 1 meter for 45 seconds in the laminar air flow. The mutant strains were compared to wild type in temperature and salinity tolerance, amylase production content and different antibiotics resistance. Results: In this study, two interesting mutant strains were isolated and named B.L.2.M.1 and B.L.2.M.2. Mutations caused many changes in bacteria such as cell growth speed and enzyme production content. Differences in cell growth, productio...
Background and Objective: Present study aimed to investigate response of metallothionein (MT) in ... more Background and Objective: Present study aimed to investigate response of metallothionein (MT) in oyster Crassostrea sp. to the experimental concentrations of Hg and Cd in order to assess the possibility of MT usage as a biomarker of Hg and Cd contamination in this mollusk. Materials and Methods: Oysters were collected from docks of Imam Khomeini Port. After seven days acclimation period in laboratory, they were exposed with Hg at concentrations of 15 and 75 μg.L-1 and Cd at concentrations of 15 and 150 μg.L-1 for 14 days. MT levels were measured through spectrophotometric method after extraction and precipitation. After preparation and acid digestion, the concentration of heavy metals was quantified by atomic absorption. Results: The amounts of MT in oysters exposed with Hg and Cd were significantly increased compared with control samples (P<0.05). The highest value of MT biosynthesis in oysters exposed with concentrations of Hg and Cd were 137.2±7.6 and 312.4±17.9 μg.g-1 w.w respectively. Exposing the oysters with Cd induced biosynthesis of MT more than three times compared with control samples. Among Hg and Cd, there was only significant correlation between biosynthesis of MT and Cd bioaccumulation in oysters (P<0.01). Conclusion: MT protein in oyster Crassostrea sp. can be considered as a suitable biomarker of Cd contamination in body and environment. Hence, it could be used for assessing and monitoring ecosystems.
Hepcidin25 is a small cysteine-rich peptide hormone known as a new class of antimicrobial peptide... more Hepcidin25 is a small cysteine-rich peptide hormone known as a new class of antimicrobial peptides. The purpose of the present study was to express, purify and investigate the antibacterial properties of recombinant human hepcidin25 protein production in Escherichia coli. Human hepcidin25 gene was optimized and fused to a small ubiquitin-related modifier (SUMO) gene for higher expression. Then SUMO-hepcidin25 was cloned into the pET-32a (+) vector and expressed in E.coli Origami. The fusion protein with a molecular weight of approximately 35 kDa was analyzed on SDS-PAGE gel. The highest expression was observed after 6 hours induction and the fusion protein consisted approximately 47 % of the total cellular protein. The purified SUMO-hepcidin25 purity was determined to be higher than 95 %, with a final yield of 3.9 mg L-1 of media. The recombinant hepcidin25 showed antibacterial activity against both gram negative (Klebsiella pneumonia) and gram positive)Staphylococcus aureus and Bacillus cereus(bacteria with minimum inhibitory concentrations (MICs) of 150 µg mL −1 , 18.7 µg/mL −1 and 37.5 µg/mL −1 , respectively. These results indicated that thioredoxin and SUMO dual fusion system is an efficient production system for synthesis functional human hepcidin25.
Background: In recent decades RNAi has become a novel and effective tool to silence gene expressi... more Background: In recent decades RNAi has become a novel and effective tool to silence gene expression. The expression of shRNAs against target genes has now become a standard and powerful strategy for antiviral therapeutic approaches. Infectious bovine rhinotracheitis (IBR) is a domestic and wild ruminant viral disease that caused by bovine herpesvirus-1. Occasionally, its entry into the herd may cause vigorous economic damage due to losing weight, reduced output and limitation on livestock international trade. Vaccines against the disease are not fully effective and the need for an effective therapeutic approach is highly felt. In the present study an RNAi-based gene therapy was designed and implemented. Due to the essential role of UL25 gene for BHV-1 replication. Method: the sequence of UL25 gene was purposed for designing shRNA molecules. The UL25 gene sequence was extracted from the NCBI database and suitable shRNA molecules were purposed by using online soft wares. Three recombinant lentiviral vectors expressing short hairpin RNAs (shRNAs) versus the UL25 gene of IBRV were constructed and co-transfected into HEK 293T cells. The effectiveness of designed shRNA molecules was assayed by calculating TCID50 titers and observation of BHV-1 cytopathic effects. Results: The nal calculations showed that all shRNAs had antiviral effects. In the TCID50 test, the shRNAs 1, 2 and 3 su ciently decreased BHV1 output in comparison with the control groups (almost 98.22%, 99.63% and 99.54%, respectively than cells with IBRV inoculation and nearly 90.84%, 97.9% and 96.84%, respectively than cells with ORFV and scrambled vector inoculation). ShRNA-2 achieved maximal inhibition of viral replication. Conclusion: The results indicated that shRNAs targeting the UL25 gene showed considerable antiviral attributes and reduced IBRV multiplication in MDBK cells. In conclusion; RNA interference can be used as a gene therapy instrument against BHV-1 virus.
International Journal of Peptide Research and Therapeutics, Aug 16, 2022
Anti-microbial peptides (AMPs) are important factors of the innate immune system which perform a ... more Anti-microbial peptides (AMPs) are important factors of the innate immune system which perform a comprehensive spectrum of biological activities. The present study aims to examine the expression, puri cation, and cytotoxic effects of recombinant PI-laterosporulin10 (LS10) a defensing-like peptide. PI is a transmembrane peptide that helps to increase penetration and speci city of peptide targeting. PI-LS10 sequence was optimized and fused to a Xa protease site to achieve higher levels of expression. Then Xa-PI-LS10 was cloned into the pET-32a (+) vector and expressed in E. coli BL21 (DE3). The fusion protein with a molecular weight of about 25.3 kDa was analyzed using the SDS-PAGE. The maximum expression was observed 8 hours post-induction. After enzymatic cleavage, the purity of PI-LS10 was >300 µg/ ml as the nal yield. The recombinant PI-LS10 showed anti-cancer characteristics against breast cancer cell lines. MCF7 (non-triple negative) and MDA-MB-231 (triple negative) cells were treated with different concentrations of the PI-LS10 in order to evaluate their cytotoxic activity and e ciency in targeting triple negative cells. Our results suggest that PI-LS10 had a signi cant cytotoxic effect against breast cancer cells in comparison to the normal cell line. Triple negative breast cancer cell (MDA-MB-231) proliferation inhibited higher in 72 hours after treatment compare MCF7 cells. In conclusion, the PI-LS10 peptide was produced successfully and its favorable anti-cancer activity against a breast cancer cell line was con rmed.
The production of natural antibiotic peptides has emerged as an important mechanism of innate imm... more The production of natural antibiotic peptides has emerged as an important mechanism of innate immunity in plants and animals. Defensins are diverse members of a large family of antimicrobial peptides, contributing to the antimicrobial action of granulocytes, mucosal host defence in the small intestine and epithelial host defence in the skin and elsewhere. This review, inspired by a spate of recent studies of defensins in human diseases and animal models, focuses on the biological function of defensins.
Background and Aims: Multicellular organisms immune system has less than 100 amino acid peptide. ... more Background and Aims: Multicellular organisms immune system has less than 100 amino acid peptide. Defensins are one of the largest family of these antimicrobial peptides and because it is applicable as much to the benefit of new generations of antibiotics. The purpose of this study was to evaluate gene expression of beta defensin 2 this is antimicrobial peptides. Methods: In this study the gene sequences BNBD2 used considering codons in E coli. and in expression vector pET-32a (+) was sub cloned. E.coli strain BL21 by the recombinant vector transfection and protein expression under IPTG induction and SDS-PAGE electrophoresis and Western blot was evaluated. Results: Recombinant proteins were expressed with IPTG. Molecular weight on gel electrophoresis (KD25) with molecular weight equal to that predicted by theoretical calculations. Techniques western blotting confirmed the presence of this protein. Conclusions: This study confirmed the sub-cloning and expression BNBD2. The high activity of antimicrobial peptides in vitro in the near future can be effective in treating bacterial infections in vivo.
Journal of the Hellenic Veterinary Medical Society, Jan 28, 2022
In silico design of short hairpin RNA (shRNA)Molecules for DNA pol gene of Contagious Ecthyma vir... more In silico design of short hairpin RNA (shRNA)Molecules for DNA pol gene of Contagious Ecthyma virus (ORFV)
Background: Border disease is believed to be one of the most important diseases in the animal hus... more Background: Border disease is believed to be one of the most important diseases in the animal husbandry industry, which has not yet been eradicated in Iran. The development of approaches based on the application of interfering RNA (RNAi) for antiviral therapy has attracted a great deal of attention over the recent years. The present research was conducted to design, construct, and apply shRNA against the NS3 gene of BDV to evaluate the prevention of BDV proliferation in the cell culture system. For this purpose, the suitable oligonucleotide sequence of NS3 gene coding was selected utilizing BDV-X818 strain. Afterwards, using shRNA design software, shRNA molecules were designed and synthesized. These shRNAs were cloned into the desired vectors and were finally transfected in HEK293T cells employing the third generation of lentiviral packaging system. Subsequently, these shRNA expressing lentiviruses were transduced to the MDBK cell line to challenge to border virus. In order to evaluate the efficacy of shRNAs, the viral infectious titer and RNA copy number were calculated with TCID50 and Real-time RT-PCR tests, respectively. Results: The results revealed that shRNAs 1, 2, and 3 decreased viral RNA by more than 90% compared to the control groups. BDV titer noticeably decreased after the challenge with shRNAs 1, 2, and 3 from ∼88% up to 99% in comparison with the control groups. Conclusions: Overall, it could be concluded that RNAi may be considered as a strong treatment proposal against viruses, such as BDV.
Background and purpose: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscul... more Background and purpose: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder. Carrier frequency studies of SMA have been reported for various populations. Although no large-scale population-based studies of SMA have been performed in Iran, previous estimates have indicated that the incidence of autosomal recessive disorder partly because of the high prevalence of consanguineous marriage is much higher in the Iranian population than in other populations. Methods: In this study, we used a reliable and highly sensitive quantitative real-time PCR assay with SYBR green I dye to detect the copy number of the SMN1 gene to determine the carrier frequency of SMA in 200 healthy unrelated, non-consanguineous couples from different part of Iran. Results: To validate the method in our samples, we determined the relative quantification (RQ) of patients with homozygous deletion (0.00) and hemyzygous carriers (0.29-0.55). The RQ in 10 of 200 normal individuals were within the carrier range of 0.31-0.57, estimating a carrier frequency of 5% in the Iranian population. Conclusions: Our data show that the SMA carrier frequency in Iran is higher than in the European population and that further programs of population carrier detection and prenatal testing should be implemented.
Jentashapir Journal of Cellular and Molecular Biology
Border disease virus (BDV) is a serious pathogen of sheep and goats worldwide. The virus causes c... more Border disease virus (BDV) is a serious pathogen of sheep and goats worldwide. The virus causes considerable economic losses in animal husbandry. Therefore, developing effective and genetic-based therapies to control viral infections, such as BDV, has been remarkable over recent years. A common way to evaluate such therapeutic strategies is by cloning the desired fragment into appropriate vectors to develop cell lines expressing it. Due to the required duties of the NS3 gene for BDV proliferation, this gene's HELICc and nucleotide binding site domain was synthesized and cloned in a lentiviral vector upstream of the GFP gene. The cloning accuracy was verified by digestion with restriction enzymes and sequencing. Thus, the BDV-NS3 HELICc and nucleotide binding site carrying plasmid was prepared successfully and will be applied in a second or third lentiviral packaging system for stable expression of the desired gene.
Journal of the Hellenic Veterinary Medical Society
Contagious ecthyma is an infectious skin disease of ruminants caused by the ORF virus (ORFV) that... more Contagious ecthyma is an infectious skin disease of ruminants caused by the ORF virus (ORFV) that is a member of genus Parapoxvirus of the Poxviridae family. In addition to the significant effects on lambs and human, ORFVs have been recently shown to infect other hosts. The disease causes significant economic damages to the sheep industry, so attempts to eliminate it must be taken into.RNA interference (RNAi) is an evolutionarily conserved mechanism in which the expression of homologous target genesis is suppressed by means of double-stranded RNA molecules. Since RNAi can be considered as a therapeutic method for viral gene silencing, we tend to make the most of this capability. The present study aims to design potential shRNAs to knockdown the DNA-polymerase gene coded by ORF025.A significant number of computational methods such as clustal omega website to target alignment, BLAST-NCBI to similarity search, CLC software to secondary structure prediction, BLOCK-iTRNAi Designer and WI...
Anti-microbial peptides (AMPs) are important factors of the innate immune system which perform a ... more Anti-microbial peptides (AMPs) are important factors of the innate immune system which perform a comprehensive spectrum of biological activities. The present study aims to examine the expression, purification, and cytotoxic effects of recombinant PI-laterosporulin10 (LS10) a defensing-like peptide. PI is a transmembrane peptide that helps to increase penetration and specificity of peptide targeting. PI-LS10 sequence was optimized and fused to a Xa protease site to achieve higher levels of expression. Then Xa-PI-LS10 was cloned into the pET-32a (+) vector and expressed in E. coli BL21 (DE3). The fusion protein with a molecular weight of about 25.3 kDa was analyzed using the SDS-PAGE. The maximum expression was observed 8 hours post-induction. After enzymatic cleavage, the purity of PI-LS10 was >300 µg/ ml as the final yield. The recombinant PI-LS10 showed anti-cancer characteristics against breast cancer cell lines. MCF7 (non-triple negative) and MDA-MB-231 (triple negative) cells...
The author regrets that, in the article titled ''Functional analysis of recombinant codon-optimiz... more The author regrets that, in the article titled ''Functional analysis of recombinant codon-optimized bovine neutrophil b-defensin", he missed to include the details of the reference in the legend of Fig. 5. The details of the missed reference is as follows: ''Karimi N, Saffar B, Ghaedi K, Mobini M. ''Optimization of expression, purification and handling anti-bacteria feature protein of bovine neutrophil beta defensin 2 in E. coli."
Background: Colorectal cancer (CRC) is one of the most common cancers in the world. High activity... more Background: Colorectal cancer (CRC) is one of the most common cancers in the world. High activity of CyclinD1 gene has been seen in the progress of this cancer. A common polymorphism (G870A) in exon 4 of CyclinD1 produces a variant transcript with longer half-life and may cause uncontrollable cellular growth thus contributing to cancer development. This study was performed to evaluate the frequency of CCND1 G870A polymorphism between CRC cases and controls. Methods: DNA samples from peripheral blood leukocytes of 50 patients with sporadic colorectal cancer and 50 healthy subjects were extracted. CCND1 G870A polymorphism was genotyped by polymerase chain reaction -restriction fragment length polymorphism (PCR-RFLP) in healthy subjects and patients. Statistical analysis was performed by chi-squared test via SPSS software. Findings: Relationship between CCND1 G870A polymorphism in allele frequencies between cases and controls were not observed (P = 0.204). While the frequency of AA gen...
Introduction:The amylase enzyme hydrolyze starch and break glycosidic internal bonds of this poly... more Introduction:The amylase enzyme hydrolyze starch and break glycosidic internal bonds of this polysaccharides. Amylases are important industrial enzymes for a wide range of applications including starch conversion to sugar syrup, dextrin production, and use in the pharmaceutical and medical industries. Materials and methods: In this study, thermophilic bacterium was isolated and purified from the Qynarcheh hot spring in Ardabil. Desired strain suspension in nutrient broth medium was placed in front of UV rays with a wavelength of 254 nm at a distance of 1 meter for 45 seconds in the laminar air flow. The mutant strains were compared to wild type in temperature and salinity tolerance, amylase production content and different antibiotics resistance. Results: In this study, two interesting mutant strains were isolated and named B.L.2.M.1 and B.L.2.M.2. Mutations caused many changes in bacteria such as cell growth speed and enzyme production content. Differences in cell growth, productio...
Background and Objective: Present study aimed to investigate response of metallothionein (MT) in ... more Background and Objective: Present study aimed to investigate response of metallothionein (MT) in oyster Crassostrea sp. to the experimental concentrations of Hg and Cd in order to assess the possibility of MT usage as a biomarker of Hg and Cd contamination in this mollusk. Materials and Methods: Oysters were collected from docks of Imam Khomeini Port. After seven days acclimation period in laboratory, they were exposed with Hg at concentrations of 15 and 75 μg.L-1 and Cd at concentrations of 15 and 150 μg.L-1 for 14 days. MT levels were measured through spectrophotometric method after extraction and precipitation. After preparation and acid digestion, the concentration of heavy metals was quantified by atomic absorption. Results: The amounts of MT in oysters exposed with Hg and Cd were significantly increased compared with control samples (P<0.05). The highest value of MT biosynthesis in oysters exposed with concentrations of Hg and Cd were 137.2±7.6 and 312.4±17.9 μg.g-1 w.w respectively. Exposing the oysters with Cd induced biosynthesis of MT more than three times compared with control samples. Among Hg and Cd, there was only significant correlation between biosynthesis of MT and Cd bioaccumulation in oysters (P<0.01). Conclusion: MT protein in oyster Crassostrea sp. can be considered as a suitable biomarker of Cd contamination in body and environment. Hence, it could be used for assessing and monitoring ecosystems.
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Papers by behnaz saffar