Showing papers in "Molecular Biology Research Communications in 2017"

Anticancer activity of cow, sheep, goat, mare, donkey and camel milks and their caseins and whey proteins and in silico comparison of the caseins.

Abstract: The present investigation was carried out to evaluate anticancer activity of cow, goat, sheep, mare, donkey and camel milks and their casein and whey proteins against MCF7 cell line. The structure-based properties of the casein proteins were also investigated, using bioinformatics tools to find explanation for their antitumor activities. The effect of different milks and their casein and whey proteins on MCF7 proliferation was measured using MTT assay at different concentrations (0.5, 1 and 2 mg/ml). The results showed that mare, donkey, cow and camel milks and their casein and whey proteins have potent cytotoxic activity against MCF7 cells in a dose dependent manner while sheep and goat milks and their proteins did not reveal any cytotoxic activity. The in silico results demonstrated that mare, donkey and camel caseins had highest positive and negative charges. The secondary structure prediction indicated that mare and donkey caseins had the maximum percentage of α helix and camel casein had the highest percentage of extended strand. This study suggests that there is a striking correlation between anti-cancer activity of milk caseins and their physicochemical properties such as alpha helix structure and positive and negative charges. In conclusion, the results indicated that mare, camel and donkey milks might be good candidates against breast cancer cells.

Genetic polymorphisms of superoxide dismutase-1 A251G and catalase C-262T with the risk of colorectal cancer

Abstract: Oxidative stress is significant in numerous types of disease including cancer. To protect cells and organs against reactive oxygen species (ROS), the body has evolved an antioxidant protection system that involved in the detoxification of ROS. Single nucleotide polymorphisms (SNP) of anti-oxidative enzymes may dramatically change the activity of the encoded proteins; therefore, certain alleles can be established as risk factors for some kind of multi-factorial diseases including cancer. In present study we investigate the possible association between polymorphisms of superoxide dismutase 1 (SOD1, OMIM: 147450) and catalase (CAT, OMIM: 115500) genes and the risk of colorectal cancer (CRC). The study included 204 colorectal cancer patients and 239 healthy control group matched for gender and age. Genotyping of SOD1 A251G and CAT C-262T were done by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method. There was no significant association between CAT C-262T polymorphism and susceptibility to CRC (P>0.05). The carries of the G allele of SOD1 significantly showed higher prevalence in CRC patients compared with the control group (OR=1.84, 95% CI=1.13-2.98, P=0.013). We assessed the effect of combination of genotypes of the study polymorphisms on the risk of CRC. We found that the combination of AG+GG (SOD1) and CC (CAT) increases the risk of developing CRC (OR=2.38, 95% CI=1.25-4.52, P=0.008).

Identification of miR-24 and miR-137 as novel candidate multiple sclerosis miRNA biomarkers using multi-staged data analysis protocol

Abstract: Many studies have investigated misregulation of miRNAs relevant to multiple sclerosis (MS) pathogenesis. Abnormal miRNAs can be used both as candidate biomarker for MS diagnosis and understanding the disease miRNA-mRNA regulatory network. In this comprehensive study, misregulated miRNAs related to MS were collected from existing literature, databases and via in silico prediction. A multi-staged data integration strategy (including the construction of miRNA-mRNA regulatory network and systematic data analysis) was conducted in order to investigate MS related miRNAs and their regulatory networks. The final outcome was a bi-layer MS related regulatory network constructed with 27 miRNAs (seven of them were novel) and 59 mRNA targets. To verify the accuracy of the bioinformatics strategy three novel and five previously reported miRNAs from the network model were selected for experimental validation using the real-time PCR assay. The obtained results proved the accuracy of the network. The expression of themiR-24 and miR-137(as novel MS candidate biomarker) and miR-16, and miR-181 (as previously reported MS candidate biomarker) showed significant deregulation in 33 MS patients compared to the control. The optimized data integration strategy conducted in this study found two miRNAs (miR-24and miR-16)that can be considered as candidate biomarkers for MS and also has the potential to generate a regulatory network to aid in further understanding the mechanisms underlying this disease.

Pharmacogenetic variation of SLC47A1 gene and metformin response in type2 diabetes patients.

Abstract: Type 2 diabetes mellitus is a worldwide epidemic disorder with considerable health and economic consequences. Metformin is one of the most commonly prescribed oral antidiabetic drugs. Pharmacogenetic studies showed that variants in genes related to the pharmacokinetics of metformin were associated with glucose-lowering effect of metformin. The aim of this study was to evaluate pharmacogenetic variation in SLC47A1 (rs2289669) and metformin response in type 2 diabetes patients. Seventy one patients with type 2 diabetes were included in the study. The genotypes were determined by Tetra–ARMS–PCR method. There was a significant association between the study polymorphism and the response to metformin treatment with the highest HbA1C reduction in AG genotype. In the dominant model for A allele (AA+AG vs GG), patients with A allele had highest HbA1C reduction in response to metformin.

Biotic elicitation for scopolamine production by hairy root cultures of Datura metel.

Abstract: The (-)-hyoscyamine, atropine and scopolamine (hyoscine) are three valuable tropane alkaloids while scopolamine is the most important member of this group for the pharmaceutical industry due to its higher demand compared to hyoscyamine and atropine. Scopolamine is an anticholinergic reagent with several therapeutic applications. In the current study, the hairy roots culture of Daturametel was used as an advantageous method for production of scopolamine. The hairy roots are formed by Agrobacterium rhizogenes and have genetic stability, high growth rate and lateral branching. In this study, the effect of Bacillus cereus and Staphylococcus aureus as biotic elicitors on the production of scopolamine in D.metel hairy roots was investigated. The amount of scopolamine in the hairy roots was detected by HPLC analysis and compared with control samples after 0, 12 and 24 hours. Results showed that, B. cereus and S. aureus enhanced scopolamine production in the culture while the atropine content was decreased. Although in the control samples with no bacterial elicitation no scopolamine was detected, elicitation by B. cereus caused production of scopolamine and about 0.03 gram and 0.017 gram of it was detected in 100 gram dried D.metel hairy roots after 12 and 24 hours respectively. In S. aureus elicited hairy roots, scopolamine was not produced after 12 hours. However, about 0.025 gram of this tropane alkaloid was detected in 100 gram dried hairy roots after 24 hours. In conclusion, S. areus and B. cereus induced the scopolamine production in D. metel hairy roots.

Molecular and phylogenetic characterization of Thelohanellus boggoti Qadri, 1962 (Cnidaria, Myxosporea, Bivalvulida) infecting the fin of Indian minor carp Labeo dero (Hamilton, 1822).

Abstract: A myxozoan parasite belonging to the genus Thelohanellus Kudo, 1933 (Cnidaria, Myxosporea, Bivalvulida) was isolated from the fins of Labeo dero inhabiting Ranjit Sagar Wetland, Punjab, India. The plasmodium was 0.5-0.7 mm in diameter each containing 80- 100 number of myxospores. Myxospores were egg shaped to ovoidal in valvular view having bluntly pointed anterior and broad rounded posterior end measuring 8.36x 4.77 µm. Polar capsule was single, broadly pyriform in shape, measuring 4.77x3.98 µm in size containing a polar filament coiled perpendicular to the longitudinal axis of myxospore body making 7-9 turns. Blast analysis of 18S rDNA sequence of the isolate demonstrated 98% homogeneity with Thelohanellus sp.(KU155561, Unpublished), followed by91% with T. caudatus (KM252684) and Thelohanellus sp. FCO (KR819273). The intensity of infection was recorded to be light as indicated by fin plasmodial index (FPI = 1). In the present study, T. boggoti has been described using 18S rRNA gene and phylogenetic analysis using MEGA.6.

Transcript analysis of some defense genes of tomato in response to host and non-host bacterial pathogens.

Abstract: The transcript levels of six defense genes including pathogenesis-related gene 1 (PR-1), pathogenesis-related gene 2 (PR-2), pathogenesis-related gene 5 (PR-5), lipoxygenase (LOX), phenylalanine ammonia-lyase (PAL) and catalase (CAT) were investigated in tomato plants inoculated with Xanthomonas axonopodis pv. phaseoli as a non-host pathogen and X. euvesicatoria as a host pathogen. Activation of all the genes was confirmed in both host and non-host treatments. Additionally, the results showed stronger expression of majority of the genes (PR-1, PR-2, LOX, PAL and CAT) in non-host treatment compared to host treatment at least at early hours after inoculation. These data suggest that faster and more expression of PR-1, PR-2, LOX, PAL and CAT might have a role in non-host resistance of tomato against X. axonopodis pv. phaseoli.

Gene variants of CYP1A1 and CYP2D6 and the risk of childhood acute lymphoblastic leukaemia; outcome of a case control study from Kashmir, India

Abstract: Studies on associations of various polymorphisms in xenobiotic metabolizing genes with different cancers including acute lymphoblastic leukaemia (ALL) are mixed and inconclusive. The current study analyzed the relationship between polymorphisms of phase I xenobiotic metabolizing enzymes, cytochromes P450 1A1 (CYP1A1) and CYP2D6 and childhood ALL in Kashmir, India. We recruited 200 confirmed ALL cases, and an equal number of controls, matched for sex, age and district of residence to the respective case. Information was obtained on various lifestyle and environmental factors in face to face interviews with the parents/attendants of each subject. Genotypes of CYP1A1 and CYP2D6 were analyzed by polymerase chain reaction and restriction fragment length polymorphism method. Logistic regression models were used to calculate odds ratios (ORs) and 95% confidence intervals (95% CIs). Compared to the GG genotype, we found a higher ALL risk in subjects who harbored variant (AA) genotype (OR=20.9; 95% CI: 6.01-73.1, P<0.0001) and AG genotype (OR=42.6; 95% CI: 8.3-217.5, P<0.0001) of CYP2D6*4 polymorphism. Although, we found a significant association of CYP1A1*2A polymorphism with ALL risk, but the risk did not persist in the adjusted model (OR=6.76; 95% CI: 0.63–71.8, P=0.100). The study indicates that unlike CYP1A1*2A, CYP2D6*4 polymorphism is associated with ALL risk. However, more replicative studies with larger sample size are needed to substantiate our findings.

Comparative genomics of human stem cell factor (scf)

Abstract: Stem cell factor (SCF) is a critical protein with key roles in the cell such as hematopoiesis, gametogenesis and melanogenesis. In the present study a comparative analysis on nucleotide sequences of SCF was performed in Humanoids using bioinformatics tools including NCBI-BLAST, MEGA6, and JBrowse. Our analysis of nucleotide sequences to find closely evolved organisms with high similarity by NCBI-BLAST tools and MEGA6 showed that human and Chimpanzee (Pan troglodytes) were placed into the same cluster. By using JBrowse, we found that SCF in Neanderthal had a single copy number similar to modern human and partly conserved nucleotide sequences. Together, the results approved the gene flow and genetics similarity of SCF among human and P. troglodytes. This may suggest that during evolution, SCF gene transferred partly intact either on the basis of sequence or function from the same ancestors to P. troglodytes, the ancient human like Neanderthal, and then to the modern human.

Association of glutathione S-transferase M1 and T1 polymorphisms and temperament.

Abstract: Mizaj (Temperament) is one of the basic concepts of Iranian Traditional Medicine (ITM), which has great importance in diagnosis and treatment of diseases, as well as maintaining the ideal healthy state of an individual. However, very little is known about the biological mechanisms of mizaj dependence treatment in practical ITM. This study was aimed to evaluate any association between the mizaj and glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) polymorphisms in healthy people. The samples included 247 healthy males from Fars province, southern Iran. The mizaj was determined using a self-reported mizaj identification scale. Subjects with equilibrium or any of four simple mizajes (warm, cold, moist, and dry) were included in the study. The GSTM1 and GSTT1 genotypes were determined using a polymerase chain reaction (PCR)-based method. No any differences in the frequency of GSTM1/T1 polymorphism between equilibrium and each of other mizaj groups were found. However, when equilibrium, moist, and dry groups were pooled and as a "medium warmness" group were compared to the warm group, the frequency of GSTT1-null in the warm group was significantly higher compare to that in the medium warmness group. Also, the combination genotypes of GSTM1 and GSTT1 was associated with the warmness; that is, individuals with combination of GSTM1 positive and GSTT1 null were more susceptible to have a warm temperament. This study indicated that the mizaj could be affected by GSTM1/T1 genes, although further research with larger samples is needed to reach full conclusions.

Immunogenic evaluation of chimeric recombinant protein against ETEC, EHEC and Shigella.

Abstract: Diarrheal diseases still remain health problem worldwide and out of many bacteria responsible for, Shigella and pathogenic Escherichia cause the most diarrheas in the world. Shigellacause bacterial dysenteries and shigellosis through invasion where the most effective proteins for pathogenesis is Ipac. Critical virulence protein for ETEC infection is CFA/I with two subunits called cfab and cfae. . Attachment of EHEC is the main step of infection and the protein Intimin plays the key role in this function. Protection against the vast majority of responsible pathogens of diarrheas requires development of the combination vaccine against Shigella, ETEC and EHEC. In the present study, a multisubunitprotein (CII) containing immunologically significant parts of CfaB, IpaC and Intimin was designed. The chimeric gene (CII) was codon optimized and analyzed with different bioinformatic servers, then synthesized and expressed in E. coli. Mice, Guinea pig and, Caco-2 Cell line were used as challenge models for EHEC, shigella and ETEC respectively. The chimeric protein induced significant immune response and therefore could be a suitable vaccine candidate against these three pathogens.

Assessment of genetic diversity of an endangered tooth-carp, Aphanius farsicus (Teleostei: Cyprinodontiformes: Cyprinodontidae) using microsatellite markers

Abstract: Genetic structure of an endemic tooth-carp fish, Aphanius farsicus from four different water bodies in the Maharlu Lake basin was investigated by applying five microsatellite markers. All of the five examined microsatellite loci showed polymor-phism pattern. A total of four alleles were detected at five microsatellite loci, with an average of 2.8 to 3.5 alleles per locus. Average values of observed and expected heterozygosity were 0.95±0.09 and 0.64±0.02 respectively. None of the tests of linkage disequilibrium were significant between each pair of loci and no deviation from Hardy-Weinberg equilibrium were detected to test for heterozygote deficiency within populations. The Nei's genetic distance values ranged between 0.03 - 0.13. Analysis of pairwise genetic differentiation between each pair of the populations revealed that fixation index (FST) values ranged from 0.013 to 0.039 and RST ranged from 0.005 to 0.065. High genetic diversity observed within the populations (99%) and low diversity (1%) among them indicating probably high level of gene flow among the studied populations of Fars tooth-carp at the present time or in the past. Regarding low genetic differentiation among the studied populations and results of population assignment test, two hypotheses are suggested and supporting evidence for each hypothesis are provided.

In silico design and expression of a novel fusion protein of HBHA and high antigenic region of FAP-P of Mycobacterium avium subsp. paratuberculosis in Pichia pastoris

Abstract: Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne's disease in ruminants and there has been a shift in the public health approach to MAP and human diseases like Crohn's disease. The prevention of infection by MAP in ruminants is thought to deter the high impact of economic losses in the level of dairy industry and possible spreading of this pathogen in dairy products. The present study was done to investigate the construction and expression of the soluble form of a novel fusion protein, consisting of Heparin-binding hemagglutinin (HBHA) and high antigenic region of Fibronectin Attachment Protein-P (FAP-P), in order to introduce as a Th1 inducer subunit vaccine against MAP. HBHA is a mycobacterial adhesin and it has been demonstrated that a HBHA-specific IFN-γ response, in latent M. tuberculosis infection, depends on the methylation of the antigen. Further, FAP-P induces Th1 polarization. Because methylation of HBHA was not performed in E. coli, Pichia pastoris was chosen as the host. The desired fusion protein had a similar 3D structure to that of HBHA with its native form and post-translational methylation in C-terminal. Hence, the uptake of the purified fusion protein will be done by M cells because ofHBHA, and cell-mediated immunity will be induced because of both antigens. Eventually, successful construction and expression of the newly-designed chimeric protein under the mentioned conditions is reported in this article.

Genetic diversity analysis of Morchella sp.by RAPD

Abstract: This study investigated the genetic diversity of morchella species using randomly amplified polymorphic DNA markers. Morchella species are an important group of edible mushrooms belonging to the family Helveliaceae with medicinal and economical significance. In this study we have developed an efficient method of genomic DNA isolation which was amplified by eight RAPD primers to test the polymorphism in three species of morchella. Out of all eight primers tested in current study, one of them (B8) resulted 100% polymorphism among the three studied species. Based on the RAPD profile a similarity matrix was generated to construct a dendrogram for phylogenetic analysis revealing the relationship among the three species of morchella.

Bioinformatic and empirical analysis of a gene encoding serine/threonine protein kinase regulated in response to chemical and biological fertilizers in two maize (Zea mays L.) cultivars.

Abstract: Molecular structure of a gene, ZmSTPK1, encoding a serine/threonine protein kinase in maize was analyzed by bioinformatic tool and its expression pattern was studied under chemical biological fertilizers. Bioinformatic analysis cleared that ZmSTPK1 is located on chromosome 10, from position 141015332 to 141017582. The full genomic sequence of the gene is 2251 bp in length and includes 2 exons. Its cDNA length is 1900 bp with a 5'-untranslated region of 311 bp and 3'-untranslated region of 341 bp, of which 1248 bp from open reading frame encoding 415 amino acid residues with a molecular weight of 46 kDa and an isoelectric point 7.2. Also, an upstream open reading frame contains 100 aa was found at -12 position from ATG initiation codon. ZmSTPK1 with a long half-life, 10 hours in Escherichia coli, and instability index of 32.25 is classified as a stable protein. A calmodulin binding domain was found in ZmSTPK1 at position from 395 to 405 in C-terminal end. The helical wheel analysis showed that the stretch of residues Ile-395 to Asp-415 has the potential to form a charged amphiphilic -helix characteristic of a calmodulin-binding region. Two P1BS-like motifs, which are present in the promoter regions of Pi starvation-induced genes, were located at positions -48 and -867 from ATG initiation codon. The expression of ZmSTPK1 responded to available phosphate, and its expression up-regulated under phosphate starvation.

ACE I/D and MMP-7 A-181G variants and the risk of end stage renal disease.

Abstract: The variants of angiotensin converting enzyme (ACE) and matrix metalloproteinases (MMPs) genes might be involved in the pathogenesis of end stage renal disease (ESRD) and hypertension. We studied the ACE insertion/deletion (I/D) and MMP-7 A-181G variants in 99 unrelated ESRD patients and 117 individuals without renal complications from Western Iran with Kurdish ethnic background. The frequency of ACE I/D variants was not significantly different between ESRD patients and controls. However, the presence of ACE D allele increased the risk of hypertension in ESRD patients by 2.14-fold (P=0.036). The MMP-7 -181 AG genotype increased the risk of ESRD by 2.04 times (P=0.026). The present study indicated the absence of an association between the ACE I/D polymorphism with the risk of ESRD. However, the ACE D allele increased the risk of hypertension in ESRD patients. Also, the present study suggests a role for MMP-7 AG genotype in the pathogenesis of ESRD.

Assessment of vocation of rifabutin and rifapentine in replace of rifampcin in drug resistance leprosy patients: a molecular simulation study.

Abstract: The emergence of drug resistance in leprosy is a major hurdle in leprosy elimination programme. Although the problem of drug resistance is presently not acute, it is important that we collect data more systematically and monitor the trend carefully so that effective measures to combat this problem can be developed. The present study aimed at the explication of cross resistance of rifabutin and rifapentine to rifampicin which would be helpful to programme managers for implementing rifabutin or rifapentine in replace of rifampicin. In this study we built 3D model of the M. leprae rpoB using Swiss Model and the modelled structure was docked with rifampicin, rifabutin and rifapentine. We established that these 3 antibiotics interact with the same binding region in the modelled rpoB of M. leprae. Thus we conclude that vocation of rifabutin and rifapentine could not be suitable in replace of rifampicin to combat with drug resistance leprosy.

Association of VNTR polymorphism of tumor necrosis factor receptor 2 (TNFRSF1B) with pulmonary tuberculosis

Abstract: This study was designed to find out the impact of the variable number of tandem repeats (VNTR) of the tumor necrosis factor receptor 2 (TNFRSF1B) on pulmonary tuberculosis (PTB) risk in an Iranian population. This case-control study was done on 159 PTB patients and 158 healthy subjects. Bi-allelic TNFRSF1B VNTR was genotyped by polymerase chain reaction. Logistic regression analysis revealed no significant association between TNFRSF1B VNTR and PTB risk (P>0.05). Our findings proposed that TNFRSF1B VNTR polymorphism is unlikely to confer susceptibility to PTB.

Taxonomic review of Argentine mackerel Scomber japonicus (Houttuyn, 1782) by phylogenetic analysis.

Abstract: Taxonomically, Argentine mackerels were first considered as Scomber japonicus marplatensis and later as Scomber japonicus Houttuyn 1782, although, in the last years, different studies have suggested that South Atlantic mackerel species belongs to Scomber colias Gmelin 1789. These latter results, incorporated in the main fish databases (FishBase and Catalog of Fishes), promoted a phylogenetic study using cytochrome c oxidase I (COI) gene sequences taken from the Barcode of Life (FISH-BOL) database. Thus, 76 sequences of S. japonicus, S. colias, S. australasicus and S. scombrus from different regions were used; including 3 from Sarda sarda as outgroup. Among S. japonicus selected sequences are those corresponding to the Argentine mackerels collected in 2007. Phylogenetic trees were obtained by neighbor joining and maximum likelihood methods and a network of haplotypes was reconstructed to analyze the relationship between species. The results showed the clear differentiation of S. australasicus, S. scombrus and S. japonicus from the Pacific while S. japonicus from Argentina was included in the S. colias group, with genetic differences corresponding to conspecific populations (0.1%). Four of the five Argentine specimens shared the same haplotype with S. colias, and none were shared with S. japonicus from the Pacific. These results suggest that the current specific name of Argentine mackerel S. japonicus should be changed to S. colias, in agreement with several genetic studies carried out with species of the genus Scomber.

Replacement of threonine-55 with glycine decreases the reduction rate of OsTrx20 by glutathione.

Abstract: Thioredoxins (Trxs) are small ubiquitous oxidoreductase proteins with two redox-active Cys residues in a conserved active site (WCG/PPC) that regulate numerous target proteins via thiol/disulfide exchanges in the cells of prokaryotes and eukaryotes. The isoforms OsTrx23 with a typical active site (WCGPC) and OsTrx20 with an atypical active site (WCTPC) are two Trx h- type isoforms in rice that were previously found to be reduced by NADPH-dependent thioredoxin reductase and GSH/Grx system, respectively. In the present work the reduction of mutants G41TOsTrx23, T55GOsTrx20, K48DOsTrx20 and T55G-K48D OsTrx20 as well as wild types OsTrx23 and OsTrx20 were tested in the reaction containing either NADPH/NTR or glutathione (GSH). The results revealed that reduction rate of T55GOsTrx20 was remarkably decreased by GSH as compared to WtOsTrx20 highlighting the critical role of Thr-55 in interaction of OsTrx20 with GSH. On the other hand a significant decrease in the reduction rate of G41TOsTrx23 was observed in reaction containing NADPH-dependent thioredoxin reductase as compared with readuction rate of WtOsTrx23. These results suggest that first residue after N-terminal active site Cys is one of the critical residue in determination of system that Trxs can be reduced in.

Evaluation of VNTR polymorphisms of dopamine transporter gene and the risk of bipolar disorder in Zahedan, southeast Iran.

Abstract: The exact role of dopamine transporter gene (DAT1) in the pathogenesis of bipolar disorder type 1 (BD) is not understood. In the present study, we aimed to evaluate the possible association between 30, 40 and 63 bp variable number tandem repeat (VNTR) polymorphisms of DAT1 gene and the risk of type 1 (BD) in a sample of Iranian population. This case-control study was performed on 152 BD patients and 153 psychiatrically healthy subjects. Genotyping of the variant was done by polymerase chain reaction method. Totally, the findings did not support an association between DAT1 VNTR polymorphisms and the risk of BD in a sample of southeast Iranian population.

Cap analog and Potato virus A HC-Pro silencing suppressor improve GFP transient expression using an infectious virus vector in Nicotiana benthamiana.

Abstract: Transient expression of proteins in plants has become a choice to facilitate recombinant protein production with its fast and easy application. On the other hand, host defensive mechanisms have been reported to reduce the efficiency of transient expression in plants. Hence, this study was designed to evaluate the effect of cap analog and Potato virus A helper component proteinase (PVA HC-Pro) on green fluorescent protein (GFP) expression efficiency. N. benthamiana leaves were inoculated with capped or un-capped RNA transcripts of a Turnip crinkle virus (TCV) construct containing a green fluorescent protein reporter gene (TCV-sGFP) in place of its coat protein (CP) ORF. PVA HC-Pro as a viral suppressor of RNA silencing was infiltrated in trans by Agrobacterium tumefaciens, increased the GFP foci diameter to six and even more cells in both capped and un capped treatments. The expression level of GFP in inoculated plants with TCV-sGFP transcript pre-infiltrated with PVA HC-Pro was 12.97-fold higher than the GFP accumulation level in pre-infiltrated leaves with empty plasmid (EP) control. Also, the yield of GFP in inoculated N. benthamiana plants with capped TCV-sGFP transcript pre-infiltrated with EP and PVA HC-Pro was 1.54 and 1.2-fold respectively, greater than the level of GFP expressed without cap analog application at 5 days post inoculation (dpi). In addition, the movement of TCV-sGFP was increased in some cells of inoculated leaves with capped transcripts. Results of this study indicated that PVA HC-Pro and mRNA capping can increase GFP expression and its cell to cell movement in N. benthamiana.