Thirteen Start Codon Targeted (SCoT), thirteen inter simple sequence repeat (ISSR) primers and eleven random amplified microsatellite polymorphism (RAMP) primer combinations were used to assess the genetic diversity among twelve Flax (Linum usitatissimum L.) genotypes that are currently active in flax breeding programs in Egypt. They successfully produced a total of 209, 177 and 143 reproducible bands, with a polymorphism percentage of 80.38%, 64.40% and 60.83%, respectively. The highest polymorphism was obtained by SCoT marker with a polymorphic information content (PIC) value of (0.23) and marker index (MI) of (2.96) compared with ISSR markers (0.20 and 1.74, respectively) and RAMP (0.16 and 1.06, respectively). Also, data showed higher SCoT values of observed alleles (Na) 1.82, the number of effective alleles (Ne) 1.51 and the effective multiplex ratio (EMR) 12.9 compared with ISSRs (1.65, 1.41 and 8.7) and RAMP (1.63, 1.34 and 6.00, respectively). On the other hand, Shanon index (I) produced by ISSR marker (0.35) showed higher variability than SCoT (0.25) and RAMP (0.30). However, dendrograms derived from SCoT, ISSR and RAMP marker data separated the 12 flax genotypes into two main clusters. Thus, SCoT marker can be used as an effective complementary method besides ISSR and RAMP for molecular characterization of flax as well as for the determination of genetic relationships between flax cultivars.
Jojoba has become a significant plant due to its precious seed oil with numerous applications. In an attempt to explore the genetic variation and the association with different traits, ISSR analysis was conducted for the selected Jojoba clones. In the present study, sixteen Jojoba plants (clones) including 1 male and 15 female clone were selected to conduct this experiment. Ten ISSR primers were used in this study. The analysis of oil content in the seeds revealed that one plant (GIADC2) contain up to 55.93% oil, which is quite high percentage. Increasing in oil content and seed weight is not always related to each other. ISSR analysis revealed 61 monomorphic bands, where, the maximum number of monomorphic bands (9) was generated by the ISSR-3, while the lowest (1) produced by the ISSR-9. However, the total number of polymorphic bands was 70. The percentage of polymorphism was ranged from 20% (ISSR-20) to 91% (ISSR-9) with average 52.6%. The dendrogram showed two main clusters; the first main cluster has grouped three Jojoba genotypes (5, 10 and 15). While the second main cluster was divided into two sub-clusters; one sub-cluster contain 10 clones (4, 6, 8, 12, 9, 14, male, 13, 1 and 11) and the other sub-cluster contained three Jojoba clones (2, 7 and 3). The association of ISSR marker with oil content revealed that ISSR-01, ISSR-08 and ISSR-19 were associated with oil content. On the hand, the association of ISSR marker with seed weight revealed that ISSR-01, ISSR-04, ISSR-05, ISSR-09, ISSR-19 and ISSR-20 primers were associated. Finally, molecular marker such as ISSR markers is important in trait marker association study for improving plant cultivars.
The present study was conducted to create new genetic variation in the hybrid population WM10 × Gemmeiza9 of bread wheat, phenotypic evaluation of F2 and F3 segregated populations for number of spikelets/spike, number of kernels/spike, number of kernels/spikelet, number of fertile spikes per plant, 1000-kernel weight and grain yield per plant and to estimate interrelationships between all possible pairs of studied traits. An exceptional transgressive segregation exceeded its parental phenotypic values in positive direction for number of spikelets/spike, number of kernels per spike and grain yield per plant was selected from F2 generation. In F3 generation, considerable high estimates of genotypic and phenotypic coefficients of variation for grain yield/plant, number of kernels/spike, number of fertile spikes/plant, 1000-kernel weight and number of spikelets/spike were observed. Positive and significant correlation coefficients between grain yield/plant and number of spikelets/spike, number of kernels/spike, number of kernels/spikelet, number of fertile spikes/plant, and 1000-kernel weight were estimated, indicating that indirect selection for these traits would be accompanied by high grain yield in this transgressed hybrid population. High positive and significant correlation coefficients occurred between number of kernels/spike and number of spikelets/spike and between number of kernels/spike and number of kernels/spikelet. These results indicated that number of spikelets/spike and number of kernels/spikelet are important in this transgressive hybrid population in the determination of number of kernels/spike.
Aroma Egyptian Jasmine rice (EJR) is one of the most important types, It is one of the high quality rice varieties all over the world, have the fragrance gene fgr. Three cytoplasmic male sterile lines, and EJR were hybridized. The three cytoplasmic male sterility Chinese lines were: Hexi41B, Yimi15B and D297 and their crosses were Hexi41B x E.Jasmine (HJR), Yimi15B x E. Jasmine (YJR) and D297 x E. Jasmine (DJR). The results showed that the male sterile lines outperform the Egyptian Jasmine rice in most of morphological trail such as; 1000 grain weight, plant height, number of panicles plant, and grain yield for plant traits. For identification of fgr gene primers were used in PCR reaction, the fgr gene was identified in the Egyptian Jasmine rice (EJR) and its hybrid lines. The gene was also digested using three different restriction enzymes (RE) such as; EcoR1, BamH1 and Hind ІІІ. By estimating the total amount of protein in the grains, the result noticed that it reached 6.91 in the Egyptian Jasmine rice (EJR) and about 6.91(grain with cover), while it reached the highest amount in DJR hybrid as crosse between the Egyptian Jasmin rice (EJR) and male sterility lines leads to fgr gene transfer enhancement and increase of protein content in rice grains which reached up to 8.3 in D297 x Egyptian Jasmine rice hybrid (EJR). The main target of this study was to determine the fragrance gene (fgr) from EJR in cytoplasmic male strile (CMS) rice after crosses between them and the EJR, that resulting in changes of charachterstics after transferring the gene to these CMS lines.
Two fiber plant species belonging to the family Asclipidaceae; Calotropis procera and pergularia daemia were chosen for the present study. Gene expression analysis of fiber quality genes (EXPA3 and EXPA4) was achieved using quantitative real-time PCR (qRT-PCR) in two plant parts (leaf and stem) in C. procera showing a high expression level of EXPA3 gene compared to the EXPA4 gene. Moreover, the expression level in stems is higher than that in leaves regardless of plant species. Detection of genes (EXPA3) was carried out by PCR reaction producing band in Calotropis procera with sizes 850bp but for EXPA4 gene PCR product in C. procera and Pergularia daemia with sizes 450bp. The obtained fragments were sequenced and phylogenetic trees based on amino acid sequences using Mega 7.0 software were accomplished. The results revealed the close relatedness of CpEXPA3 understudy to C. procera (ABO30977.1) was recorded in the National Center for Biotechnology Information (NCBI) database. In addition, the close relatedness of CpEXPA4 understudy to CpEXPA4 in C. procera (EF434784.2) was recorded in the NCBI database. The present study recommends conducting more researches on fiber crops due to their economic importance. Furthermore, their genetic potentialities which enable them to be good genetic resources and alternatives to economically important fiber crops. Moreover, For Guidance to domestication and cultivation of inexpensive fiber resources that are tolerant to various environmental stresses and do not exhaust the farmlands.
Bagasse is a major source of lignocellulose that is produced in a huge amount in Egypt. It can be used as a second-generation biofuel starting material, fibers for paper, and substrate for high-value microbial products. The present study describes the isolation of cellulose-degrading bacterial strain, AGE-B8, isolated from rotted sugar cane bagasse. AGE-B8 bacterium was tested for the cellulases induction under various parameters such as pH, incubation temperature, incubation time, carbon sources. The best filter-paper enzyme (FPase) activity was obtained after incubation for 7 days at 34oC, while best carboxymethyl cellulase (CMCase) activity was obtained after incubation for 7 days at 37oC. The strain was identified using multilocus sequence analysis as Bacillus altitudinis. A draft assembly of the AGE-B8 genome was performed. The assembly was submitted to NCBI under project number PRJNA675729, BioSample number SAMN16712403, and SRA accession number SRR13084424. A total of 3,540 genes were identified in the produced bacterial genome assembly. Gene annotation and genetic variation analysis was performed for the identified isolate. A detailed list of all mutations, in reference to the NCBI Bacillus altitudinis reference genome, was identified. The results will significantly contribute to a better understanding of the genetics of the lignocellulosic degrading bacteria.
phylogenetic relationships of eleven accessions of (Hordeum vulgare L.) collected from different region of Egypt were assessed. Fifteen quantitative morphological traits were used, the measured data were evaluated statistically using ANOVA, phylogenetic tree were constructed using UPGMA. Also Inter simple sequence repeat (ISSR) molecular marker technique were used for DNA fingerprinting and assessing genetic diversity and phylogenetic relationships in barley germplasm. The results showed that ISSR primers produced 140 bands their size ranged between 110-1600 bp with 39.8% polymorphism percentage. Polymorphic information content PIC was 0.74 for ISSR. UPGMA dendrogram was divided into two clusters by morphological traits and ISSR analysis. Genetic similarity matrix was examined with Jacard’s coefficient, maximum similarity was found between B8 and B7 (98%) with morphological analysis both from (North Sinai) and between B9 and B10 (96%) with ISSR analysis. Determination of genetic diversity between barley is of major importance for characterization of barley germplasm, breeding programs and conservation purposes. Morphological traits and ISSR analysis are effective tools for detecting genetic variations. The results showed that H. vulgare have high ratio of variation. This study may be considered as reference study for further studies on H. vulgare and may contribute to species concept and breeding programs.
Genetic studies have been performed to distinguish two plants inhabiting a natural valley renowned as Wadi Hagul of Egypt throwing karyotype and DNA barcode analysis. The two plants were morphologically identified as Echinops spinosus and Fagonia mollis. As a result, the karyotype approach exhibited a significant variant among the certain chromosomal features involving chromosomal number, area of chromosomes, chromosomal length, arm ratio, the position of centromeres, and centromeric index. The chromosomal number of both plants was 2n=18, while the DNA barcode identified their species as E. spinosus of the family Asteraceae and F. mollis of the family Zygophyllaceae based on rbcL phylogenetic analysis. Results indicated that, F. mollis evolutionary tree was rooted into two clads, belonging to family Zygophyllaceae and supported to Fagonia latistipulata (bootstrap support of 93%). Indeed, analysis of rbcL sequence will permit to evaluate the taxonomy and systematic of different plants recovered with a good performance in clarifying genetic diversity within and between populations in the studied plants. In particular, Echinops spinosus and Fagonia mollis showed a minor divergent from the common species, therefore an urgent survey for these genotypes and their conservation is required.
Strawberry (Fragaria x ananassa) is one of the favorite fruit worldwide due to its wide health benefits, and distinct flavor and aroma. FaWRKY1 gene was suggested as a significant element intermediate defense response against various pathogens attack in strawberry. Because of the influencing role of WRKY family involving in defense network, it has become a favorable candidate for improving crops quality. WRKY can precisely recognition and binding to the down-stream promoters of transcription factors activating defense cascades. The ability of FaWRKY to enhance resistance against Macrophomina phasiolena was investigated by performing Agrobacterium-mediated transformation protocol for transient overexpression of FaWRKY1 gene in strawberry leaves (Fragaria x ananassa cv. Camarosa) to evaluate its function upon the fungal infection. However, MtNOOT gene were transiently hetero-expressed in strawberry leaves separately and in accomplishment with the FaWRKY1. We demonstrated that the existing of W-box sequences within the MtNOOT (NPR1-like gene) promoter region, which are recognized definitely by SA-induced FaWRKY DNA binding protein, increased strawberry resistance activity when the two genes are transformed in combined to each other. The severity of leaf injury was observed at three, five, and seven days post pathogen inoculation on FaWRKY1, MtNOOT, and FaWRKY1 & MtNOOT combination-transformed strawberry plants compared to the control untransformed infected plants as positive control and a healthy untransformed non-infected plants as negative control. Susceptibility to fungal infection was obviously detected and showing that the two genes combination (FaWRKY & MtNOOT) revealed the best resistance against the pathogen fungal attack followed by MtNOOT and finally by the FaWRKY1 transcription factor. Our results evidence that FaWRKY1 gene acts upstream of the heterologous MtNOOT (NPR1-like gene) and positively regulates its expression throughout plant defense activation during pathogen attack.
Successful conversion of lignocellulose waste to fermentable sugars should open the door for the production of many desired metabolites. Fungi are the best cellulose decomposers in nature. The present study describes the isolation of cellulose-degrading fungal strain, AGE-1, from cow manure. AGE-1 fungal strain was tested for the cellulases production under various pHs, incubation temperatures, incubation time, and carbon sources. The AGE-1 isolate showed the maximum activity for CMCase of 1.44 U/ml ± 0.02 on day 7 when the fungal isolate was incubated at 30 °C. The maximum activity for FPase was on day 7 and it reaches 0.88 U/ml ± 0.03 when the isolate incubated at 30oC. The strain was molecularly identified using the 18S rRNA gene and ITS as Aspergillus terreus. The strain was tested for cellulases production using solidstate fermentation and showed maximum expression of FPase and CMCase at 30oC and pH7. Various agricultural wastes were tested as substrates for cellulase production by solid-state fermentation, and sugarcane bagasse (SCB) was the best sole source of carbon for cellulase production followed by wheat straw. The isolated strain can be used as a starting step for degrading agricultural waste that can be used for the production of biofuel and the production of desired metabolites from cellulose.
In this study, a recombinant baculovirus expressing the US9 tegument protein of the Egyptian BoHV-1.1 Abu-Hammad strain (vAc-Us9) was constructed using Bac-to-Bac expression system. A coding fragment of 435bp of Us9 gene was cloned into pFastBac-1 expression vector then transferred into the baculovirus expression vector system (BEVs) for expression f BoHV-1 US9 tegument protein. The PCR analysis of the recombinant virus confirmed the successful transposition of the expression cassette into the recombinant bacmid. The expressed US9 protein in infected cell culture was assayed using Indirect Immunofluorescence Assay (IFA) in addition to SDS-PAGE and western blot using crude cell lysate. The US9 protein expressed in infected cell culture supernatant as well as cell lysate were used as a coating antigen in an indirect enzyme linked immunosorbent assay (ELISA) to examine its sensitivity for the detection of the specific antibodies that generated against BoHV-1. The effectiveness of using of the expressed US9 protein as antigen for anti-US9 detection in serum samples infected with BoHV-1 was discussed in details.
Rice (Oryza sativa L.) is a semiaquatic plant, grow well in tropical, subtropical and temperate regions and it is highly affected by water shortage. Differential display reverse transcription (DDRT) technique was used to investigate differential gene expression between two Egyptian Oryza sativa cultivars under drought treatment, in an attempt to identify stress genes whose expression is regulated by drought and define their function. Fifty-six and fourty-seven fragments were differentially expressed under drought in G179 and Sk101, respectively. These differentially display (DD-PCR) fragments were categorized into up- and down- regulated fragments. DNA sequences of 17 fragments out of the total DD-PCR fragments was subjected to a nucleotide and amino acid sequence homology search through BLAST analysis programs from the National Center for Biotechnology and Information (NCBI). Four of the DD-PCR fragments were found to have the same base sequences showed homology with chaperone proteins DnaJ15. Moreover, another three fragments with the same sequence illustrated a homology with re-trotransposon protein, putativeTy3-gypsy subclass. Narrowing down the number of DD-PCR fragments to 12 fragments. Sequence alignment analysis identified four fragment sequences with significant homology to ribosome biogenesis protein WDR12, transposon protein, putative, CACTA, En/Spm sub-class, cytokinin riboside 5'-monophosphate phosphoribo-hydrolase LOGL10 and peroxisome biogenesis protein 12 of Oryza sativa. Real time PCR for three genes cytokinin riboside 5'-monophosphate phosphoribohy-drolase LOGL10, peroxisome biogenesis protein 12 and retrotransposon protein, putativeTy3-gypsy subclass were done to confirm their concentration.
Genus Origanum is one of the most species-rich as medicinal plants and pharmaceutically in the family Lamiaceae for several multipurpose used, and thus it is an endangered plant that needs a strict conservation plan. This genus contains many plants with medical uses, and thus an objective identification method is urgently needed. DNA barcoding is a sample fast technique at molecular level in the field of identification, authentication, classification and differentiation between two Origanum species and with other species. The current investigation was conducted to identify, discriminate and authenticate Origanum vulgare L. and Origanum syriacum L. subsp. sinaicum using two chloroplast genes (coding sequences) as the most common DNA barcodes, ribulose 1, 5-biphosphate carboxylase large subunit (rbcL) and maturase K (matK) genes. The partial sequence length of rbcL gene of two Origanum species were 528bp and Lamiaceae and take them away from five plant species from different families as out group of the family Lamiaceae. The obtained results revealed that rbcL and matK genes nucleotide sequence isolated from the two Origanum species in this investi- gation showed high similarities and closely related to NCBI recoded 17 genera be- longing to the family Lamiaceae. Furthermore, a phylogenetic tree analyses were constructed based on amino acid sequence of 60 rbcL and 60 matK genes using MEGA 7 program by Maximum Likelihood (ML) method with the highest log likelihood (-2836.15) for rbcL gene and (-5692.60) for matK gene
PKR is a p53 target gene and acts an imperative role in the tumor-suppressor role of p53. Initiation of p53 by genotoxic tension prompts a significant level of PKR expression, that ensuing in translational embarrassment and cell apoptosis. The current study aims to evaluate prognostic influence of PKR gene expression in chronic HCV patients; Correlate PKR and P53 gene expression to liver function tests and also shows the role of PKR in patients infected with HCV. We concluded that PKR was established to be an independent prognostic issue indicating the vital biological significance of this gene in the HCV disease process. In spite of the restriction of this study related to sample size, it paved the way for further future studies using more samples. Further investigations on a larger scale via well-standardized performances and more samples are recommended to validate these results of ROC curve or define a suitable one.
Ethidium bromide (EtBr) is a nucleic acid intercalating agent used extensively as a fluorescent dye in molecular genetics laboratories. The current study aimed to determine the potential histo- and genotoxicity of EtBr and investigate the antigenotoxic effect of probiotic bacteria (Lactic Acid bacteria; LAB) on mammalian tissue (albino mice). Mice were randomly divided into seven groups, with seven different treatments. Different EtBr doses were used individually as drinking solutions with and without probiotic bacteria, which was introduced as a single dose as a food additive. After one month of the treatments, the liver was tested using histological assay, DNA fragmentation analysis, and quantitative RT-PCR technique. No significant genotoxic effect for EtBr on the liver was observed on histo- logical examination and DNA fragmentation analysis. However, a considerable increase in the expression of the p53 gene and correlated with the dosages increase. However, the p53 expression was altered upon applying the probiotics, while some histological changes were detected, but no DNA fragmentation was detected. The seriousness of EtBr on the organism's health is conditional with the applied doses. Probiotics were not luckier than EtBr; it did not provide expected health benefits. This unexpected action of probiotics may be due to the used dose and how to use it. Further analysis of both histological and molecular aspects using multiple controls and integrative experiments is required to explain probiotics' paradox effect versus the EtBr effect on mammalian cells and tissues. Keywords: EtBr, Probiotic bacteria, genotoxicity, q-PCR, DNA fragmentation.
Thirty-seven endophytic bacterial isolates were isolated from tissues of four salt-tolerant plant species (Phragmites australis, Tamarix nilotica, Juncus rigidus, and Halocnemum strobilaceum) growing on shores of the El-Hamra Oasis (hypersaline soda lakes, with salinity level, reaches up to 5.0 M NaCl), EL-Natrun Valley, Egypt. Isolates were determined by different morphological, physiological, and molecular characters. Sequencing data of 16S rRNA declare that the 37 isolates belong to 3 genera Bacillus, Staphylococcus, and Vibrio (accessions no. Ky608807 - KY608843). All isolates produced Indol- 3-acetic acid (IAA) when allowed to grow in LB media fortified with 0.1% L- tryptophan. The highest five potent isolates IAA- producer were selected and used to inoculate tomato (Lycopersicon esculentum) plants under greenhouse conditions. Based on plant growth characteristics that were measured at 37, 51, and 65 days post-germination; we found that 10 or 50 ml (1 x 107 CFU/ml) direct addition of bacterial culture to soil surface caused a significant increase in tomato plants' height (above-ground fresh and dry weight) compared to direct foliage spraying. When pooled together, all bacterial treatment caused a significant increase in tomato height (ranging from 4.9 up to 25.8%), fresh weight (13.2 up to 43.4%), and dry weight (42.37 up to 51.58%, with an average of 46.73%), compared to non-bacterial inoculated controls (non-treated, or IAA-sprayed) 65 days post-germination.
Precise identification of jojoba plant species using DNA barcode is very important because of the economic value of its seed oil. In this study, we selected 16 jojoba trees (clones) including one male and 15 female trees cultivated in the same farm to conduct this experiment. DNA samples were extracted from leaf tissues and subjected to PCR amplification using specific primers for rbcL gene fragment. The amplification was successful for all tested samples producing the same fragment with a size about 550 bp. Sequence analysis was performed for the purified products and consequently subjected to GenBank database analysis. The data analysis produced by BLASTn database revealed that the similarity search was from 98-100% with rbcL gene region form available DNA sequence in GenBank. Multiple sequence alignment was performed after confirmation of the DNA sequence. The alignment clearly indicated the high similarity between all DNA sequences for the tested samples. Furthermore, a phylogenetic tree was produced from the alignment process which revealed the close relationship between the tested samples. The tree can be divided into two main branches one containing samples 1-13 and 16 while the other branch contains samples 14 and 15. These results indicate that the rbcL region is a good option for molecular identification of jojoba species. Finally, the obtained sequence data were submitted to GenBank to provide the accession numbers for the nucleotide sequences.
Nowadays, Stevia rebaudiana leaves have attracted economic interest as a natural non-nutritive, non-toxic, high-intensity sweeteners as well as its therapeutic benefits. The aim of the study was established an efficient regeneration protocol to improve stevia's sweetness with enhancing productivity of stevia leaves as well as to evaluate the possible suitability of this technique to be applied to rapid production. The young leaves and hypocotyl of Stevia rebaudiana namely Spanti explants were cultured on a MS medium supplemented with different concentrations and combination of NAA, 2,4-D and BAP as plant growth regulators. The results showed that leaf explants obtained highest callus formation (100%) on MS medium supplemented with 0.5 mg/l NAA while callus formation from hypocotyl explants was (92.5%) on MS medium contains 1.0 mg/l 2,4-D and 2.0 mg/l NAA. Callus derived hypocotyl failed to differentiate, while callus resulting from young leaf explants were differentiated into shoots on MS medium supplemented with 2.5 mg/l BAP. MS containing 1 mg/l BAP and1 mg/l 2,4D showed best results in steviol glycoside components and high rebaudioside A (RA) (24.22 mg/g callus) yield comparing with other used media under inoculation for 18 hrs light. Also this media gave the high rate of RA over stevioside (1.33) as an indicator of leaves quality with the sweetness intensity rate under the same experimental conditions. Medium with 1.0 mg/l IBA was suitable for rooting stevia shoots. Plantlets were acclimatized by 64% in the soil contain beat moss, sand and perlite (1:1:1) under controlled chamber at 28°C for one month which transferred to the green house.Key words: Non-nutritive, callus formation, hypocotyl explants, steviol glycoside.
Phosphorus (P) is one of the most important plant nutrients that greatly affect overall plant growth. P-mineral always presents in soil with the insoluble condition. Phosphate compounds generally contain aluminum (Al), iron (Fe), manganese (Mn), and calcium (Ca) which vary according to soil type. Phosphate solubilizing fungi play a major role in rising soil phosphate bioavailability for plants by realizing phosphate from its compounds. The present study was aimed to isolate and characterize phosphate solubilizing fungi from Egyptian soil using a solid Pikovskaya (PVK) medium. In total, 6 fungal isolates were able to grow on Pikovskaya (PVK) medium but only one isolate (AG-A01) showed phosphate- solubilizing capacity. DNA was extracted from the isolate followed by amplification of selected Internal Transcribed Spacer (ITS) region and DNA sequencing. After analyzing the DNA sequence, the isolate AG-A01 was identified as Aspergillus niger. The fungal isolate also displayed an antagonism effect against two plant pathogens (Fusarium solani and Fusarium verticillioides). Thus, we consider this fungal isolate as a promising tool for the development of an efficient bio-fertilizer for the plant which has the potential to protect plants from pathogens especially fusarium species.
Recently, micro RNAs have been shown to regulate gene expression of genes in many organisms. They bind to target mRNA transcripts. In a sequence specific manner, including mRNA degradation, translational repression or endonucleolytic cleavage. Some studies showed that miRNA expression correlates with various cancers. Combination of multiple properties e.g. free-energy, sequence pattern, hairpin shaped precursor rather than miRNAs complementarity to their targets provide a more desirable accuracy in miRNA target prediction. Different programs were used to predict novel targets for differentially expressed miRNA in cancers tissues. Luciferase assay for PEX5, GHR and AXIN2 showed a significant decrease in luciferase production in the presence of both miR-195 and PEX5-3'UTR, GHR-3'YTR and AXIN2-3'UTR. The results emphasize that both PEX5, GHR or AXIN2 are real targets for miR-195. This confirms and verifies the prediction method that were used on this study and our previous studies. These genes are suggested to have a major role in regulation of Wnt signaling pathway, epithelial-mesenchymal transition, AKT Signaling Pathway, peroxisomal protein import and cell longevity.
