نشریه زیست فناوری دانشگاه تربیت مدرس
سال نهم شماره 1 (پیاپی 16، زمستان 1396)

The induction of artificial over-expression of miRNAs is an appropriate approach to more effective cell differentiation. The significant role of microRNA-1(miR-1) has been reported in the development and differentiation of cardiac cells. Lentivirus is an effective vector for stable cell line production. The aim of this study was the production of recombinant HEK293T with miR-1 overexpression as a biological model for cardiac studies. In this experimental study, HEK 293T cells were cultured in DMEM medium with 10% Fetal Bovine Serum (FBS) and L-glutamine 2mM and Penicillin-Streptomycin 1X in incubator medium. After cloning of miR-1 gene, recombinant clones were selected and the recombination was confirmed by sequencing. The miR-1 carrying vector and auxiliary vectors were packaged in the HEK293T to produce the recombinant virus. The infection of HEK293T by recombinant virus was performed in order to achieve stable cell line. Then, GFP fluorescent marker evaluated the efficiency of transfection and effective virus dilution. Finally, the alteration in expression level of miR-1 was assessed by qPCR. Data analysis was performed by comparing the threshold cycle and Pfaffl method. The most GFP expression was detected in transfected cells by 150 micromole dilution. GFP fluorescent marker facilitated optimization and purification of recombinant cells. qPCR investigation demonstrated the significant increase in expression of miR-1 in transfected cells in comparison to controls. The stable recombinant HEK293T miR-1 over-expressing cell line in lentivirus can be utilized as a suitable biological model for investigation of cardiac evolution and development processes.

Heavy metals are one of the most important pollutants in earth and water environments due to long-term durability. The aim of this study was to isolate phosphate solubilizing bacteria from metal waste, investigate the amount of resistance, remove the metal by it and the effect of phosphatase on removal of metals. In this experimental study, the isolation of phosphate solubilizing bacteria and detection of isolates were carried out, using biochemical and molecular tests. The phosphatase was measured by colorimetric method, the resistance of the separated to the metals with the minimum inhibitory concentration (MIC50), minimum bactericidal concentration (MBC) and the rate of removal of metals by atomic absorption was measured. The surface changes of the exposed metal cells were investigated by Fourier Transform Infrared Spectroscopy (FTIR) and the effect of phosphatase on metal removal. Data analysis was done with Duncan's test, using Excel 2013 and SPSS 20 software. Serratia proteamaculans was identified as producer of the acid phosphatase. The highest MIC and MBC were obtained for Nickel (Ni) and Lead (Pb), respectively. The most metal removal was for Pb. MIC50 of Chrome and Cadmium were obtained less than 0.1mM and 1mM, and their removal percentage by the isolate were 18% and 48%, respectively. According to the FTIR, 988.339cm-1 wavelength was observed in the cells treated by 5mM Pb that is related to the Pb3(PO4)2. The isolate showed the highest resistance and removal of Pb. The mechanism of Ni removal was associated to the cell surface, while Pb was removed by both of the cells and supernatant containing phosphatase. Serratia proteamaculans is the phosphate solubilizing bacterium in metal waste. This bacterium produces an enzyme called phosphatase, which is a cause of lead removal.

Today, due to the advent of drug resistance in cancer cells against conventional drugs, attention has been paid to the development of anti-cancer drugs with new mechanisms. Pardaxin is an amphipathic polypeptide neurotoxin.The aim of this study was to investigate the interaction of antimicrobial peptide pardaxin with DPPC (composed of 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine) bilayers by molecular dynamics simulation. In the present study, simulations for different membrane environments were designed under neutral pH conditions. At first, the Linux system was used to install the VMD 1.8.6 (Visual Molecular Dynamics) software; then, Gromacs 4.5.5 software was used to perform all the simulations. The pdb peptide structure (1XC0) was prepared from the Protein Data Bank and DPPC lipid bilayer was used for lipid-peptide simulation. During the 500 nanoseconds of simulation, the peptide was infiltrated into the membrane. In the DPPC system, at first, the number of hydrogen bonds between the peptide and the lipid bilayer were increased and, then, remained almost constant until the end of the simulation and decreased over time with the number of hydrogen bonds between peptides and water. Pardaxin contacted with the membrane surface and entered into the membrane. In the presence of the peptide, the thickness of the membrane and the range of each lipid decreased and the membrane penetration increased. The mechanism of Pardaxin is dependent on the bilayer composition, so that the pardaxin peptide contacts with DPPC lipid membrane surface and enters into it.

Bioproduction methods of nanoparticles are preferrabale to chemical and physical methods because of low energy and time expenditure. The aim of this study was to investigate the biological preparation of silver nanoparticles, using Artemisia sieberi. In this experimental study, the extract of Artemisia sieberihas was used to produce silver nanoparticles by a simple, non-toxic, and low-cost method. Formation of silver nanoparticles was established despite the presence of an absorption peak at 490nm, using spectrophotometer. The size and shape of silver nanoparticles were shown using scanning electron microscopy. Precise size and change range of nanoparticles were measured by Particle Size Analysis (PSA). FT-IR results also indicated the role of different functional groups in the synthetic process. The change in the color of the extract from pale yellow to light brown and absorption peak at about 490nm showed production of silver nanoparticles. The silver nanoparticles were mainly spherical and their diameter was in the range of 27nm to 65nm, and in some regions, they were stacked or scattered together. The mean size of nanoparticles was 70nm and the dispersion of nanoparticles was in the range of 40nm to 140nm. The silver nanoparticles derived from the Artemisia are spherical and their mean size is about 70nm. Their dispersion is between 40nm and 140nm.

Carotenoids are a vast group of lipid-soluble pigments, which are produced by variety of microorganisms. The aim of this study was to compare the production of carotenoid pigments by prokaryotic isolates of Iranian saline ecosystems and identify superior isolate. In this the experimental study, isolates were purified by culture-based methods and carotenoid extracts were analyzed by spectrophotometry in wavelength region of 400nm to 600nm. The total carotenoid content was estimated by spectrophotometry at λmax (490nm). Identity of bands was detremined by purification of bands by Thin Layer Chromatography (TLC) and analysis by High Pressure Liquid Chromatography (HPLC) and Fourier Transform Infrared Spectroscopy (FTIR). Fourty-three isolates were obtained. Eight isolates were halotolerant bacteria, 8 isolates were moderately halophile, and 27 isolates were extremely halophile. All of the strains were capable of producing carotenoid compounds. Isolate M24 with 2054μg/g production was selected as superior isolate. Thin layer chromatography exhibited 6 colored bands in colored extract of this strain and the most concentrated band was purified. After purification by TLC and HPLC, spectrophotometry in UV range showed two pics at 530nm and 465nm as the highest absorbances, which were similar to UV absorbance of α-bacterioruberin. Phylogenetic analysis of 16S rRNA gene sequence of strain M24 showed that this strain had 98% similarity with Haloarcula amylolytica BD-3. From Iranian Saline Ecosystems, 43 isolates are obtained. Eight isolates are halotolerant bacteria, 8 isolates are moderately halophile, and 27 isolates are extremely halophile. All of the isolates are capable of producing carotenoid compounds. Strain M24 is superior isolate, having 98% similarity with Haloarcula amylolytica BD-3.

The chlorinated organic compounds are the most dangerous water pollutants in industrial sites. The aim of this study was to investigate the biodechlorination of chlorinated aliphatic compounds; trichloroethylene, dichloromethane, and 1,2- dichloroethane in aqueous solution, using aerobic Sphingopyxix ummariensis bacteria. In this experimental study, aliphatic chlorinated compounds; diclormethan, trichlorethylene, and 1,2-dichloroethane with purity of 99.9% were used. A visible-ultraviolet spectroscopy was used to determine the cell growth from measuring the turbidity of the medium at 600nm. The amount of released chloride was measured by an Ion Selective Electrode (ISE). The live bacterial sample was inoculated into the Nutrient Broth medium and was incubated at 30°C and 150rpm for 24 hours. The rate of dechlorination of diclormethan, trichlorethylene, and 1,2-dichloroethane by Sphingopyxis ummariensis were measured as 1.3, 1.05, and 0.63mg/l.h, respectively. The addition of glucose and yeast extract, as co-substrate, led to an increase in the cell growth and dechlorination rate up to 3.28, 1.67 and 0.90mg/l.h, respectively. During experiment, the highest dechlorination was measured at concentration of 2.5mM, at exponential growth phase. Sphingopyxix ummariensis bacteria is capable of biodechlorination of chlorinated aliphatic compounds and can grows on trichloroethylene, dichloromethane, and 1,2-dichloroethane as a single carbon source and can decolorize them. This strain has the highest growth and removal efficiency in eliminating dichloromethane as the sole source of carbon along with glucose and yeast extract as co-substrate.

The Mesd is a universal inhibitor and has therapeutic effect against triple negative breast cancer. The peptide derived from carboxyl terminal, similar to protein, acts as an inhibitor of the pathway. The aim of this study was to investigate the probable binding sites of Mesd and its peptide derived from carboxyl terminal on LRP6 first and second beta-propeller domains from a structural point of view in drug design. This experimental study was conducted, using blind and site-directed molecular docking simulation with ClusPro and Haddock and molecular dynamic simulation. The binding sites of Mesd and the peptide on the first and second beta-propeller domains of receptor LRP6 were investigated and the selected complexes were structurally analyzed. Extensive levels of Mesd protein were found to interact with LRP6 and the levels involved in the peptide were much lower. The binding region of Mesd to LRP6 was from the carboxyl terminal. The binding region of the peptide and the protein on LRP6 was a similar region between First and Second Beta-Propeller Domains of LRP6. The RMSD and RMSF chart of the Mesd complex and its peptide was approximately the same with the first functional domain of the LRP6 co-receptor. The binding region of the peptide and the protein on LRP6 is not completely similar, but according to molecular simulation of selected complexes, the pattern of the inhibition mechanism is common and emphasizes on inter domain motion control from a structural point of view. Interactive region of each ligand is similar to a region of the co-receptor, which has maximum flexibility. Molecular docking simulation of Mesd and co-receptor shows important role of carboxyl terminal of the protein to bind to LRP6.

Age-Related Macular Degeneration (AMD) is one of the biggest causes of vision loss after 50 years of age in the world. AMD disease destroys the retinal pigment cells. Retinal tissue engineering provides a suitable environment for the growth of retinal pigment epithelium cells using different scaffolds. These scaffolds may cause interior pressure changes in eyes and thus, causes disease of the separation of pigment and retinal epithelial cells. Therefore, the purpose of this study was to simulate gelatin, gelatin-chitosan and poly-caprolactone scaffolds in the retina and compare the pressure gradient and the effect of thickness on the pressure gradient. In the present experimental study, in the first stage, three gelatin, gelatin-chitosan and poly-caprolactone scaffolds were simulated to examine the average scaffold pressure using COMSOL 5.1.1 software and Darcy law. In the next step, a gelatin-chitosan scaffold with thicknesses of 10 and 20 micron was simulated with Darcy law, to examine the effect of thickness on average pressure. The output pressure of the gelatin scaffold was calculated as 308.800Pa Which was less than the pressure level of the caroid layer And it was less than the output pressure of other scaffolds. The average pressure of gelatin-chitosan scaffold with thicknesses of 10 and 20 micron was 1997.31 and 2003.13 respectively in the last step. The gelatin scaffold produces a moderate lower pressure than the gelatin-chitosan scaffold and poly-caprolactone in the retina and it is more suitable than other scaffolds. In the simulation of gelatin-chitosan scaffold, increasing the thickness causes increased pressure and retinal impairment.

Marine macroalgae are diverse organisms with adaptation for live in stressful environments. The aim of this study was to investigate the biological activities of organic extract; n-Hexane (nH), ethylacetate (E) and methanol (M) of three green alga from family Ulvaceae, Ulva clathrata, Ulva linza and Ulva intestinalis, collected from the coast of Bandar Abbas. In this experimental study, for identification the superior species, the tested activities included antioxidant assay at gradient concentrations by ferric reducing power assay, total antioxidant capacity, total phenolic content, and brine shrimp cytotoxicity activity of these extracts on model organism, Artemia salina. Data analysis was performed by one-way analysis of variance and Duncan's multiple tests at 5% probability level using SPSS 21 software and drawing charts using Excel 2013 software. The more effective algal extracts by maximum antioxidant capacity, were recorded for M extracts of U.intestinalis, E and M extracts of U.linza and U.clathrata. The algal extract exhibited a higher antioxidant activity in comparing to ascorbic acid (as a standard) with significant differences between the extract in different concentrations (p≤0.05). The result showed the highest content of total phenol were recorded for the M extracts of U.linza and U.clathrata which confirmed the findings of other researchers that the increase in free radical scavenging activity of natural extracts is associated with the content of phenolic compounds. The highest brine shrimp cytotoxicity activity was recorded for the nH extracts of U. linza (LC50= 300.78 mg/ml). According to the results, in general, U.linza can be introduced as a priority species for biological properties and in further studies. Three green alga from family Ulvaceae, Ulva clathrata, Ulva linza and Ulva intestinalis, have antioxidant and cytotoxic activity. U.linza due to the high amount of phenol and high antioxidant power can be introduced as a priority species for biological properties.

Nylon or polyamide is one of the most used and most important polymers used in the plastic and fiber industries of the world. For this reason, its use is less sensitive to the properties of its very poor biodegradability. Therefore, the aim of the present study was the biodegradability modification of synthetic polyamide 6 (pa6) fibers via in-situ melt blending with recycled poly (lactic) acid plastic food container flakes (r-PLA) during the melt spinning process.

In this experimental study, polyamide chips 6 in textile industry and Poly (Lactic) Acid Plastic Disposable Container Flakes were used. The weight loss, mechanical properties, and surface morphology variations of pure and modified fiber samples after soil burial test were analyzed for comprehensive biodegradability study of the modified fiber samples. Data were analyzed by One-Way Analysis of Variance.

The mechanical tests performed on Norris fiber showed successful production of blend fibers with the percentages of 5, 10, 20, 30, and 40 of the components of r-PLA and A 50% r-PLA fiber sample did not have acceptable mechanical properties. The changes of PA6/r-PLA blended fibers with a significant increase in r-PLA component in the PA6 substrate were significant.

The blend modified of PA6 and Poly (Lactic) recycled samples, with a composition containing from 5% to 40% of the dispersed recycled poly-lactic acid fraction have successfully melt spinning capability. By increasing the percentage of recycled poly lactic acid in the blended fibers, the mechanical properties show improvement in samples of 5% and 10% by weight and show reduction in higher percentages. Iincreasing the biodegradability of modified PA 6 fibers with increasing the r-PLA content is obviously confirmed.

Growth hormone is a non-glycosylated polypeptide strand of the pituitary glands of all vertebrates that has a wide range of biological activities and considering the importance of this hormone and its importance and diverse therapeutic applications in medicine, its recombinant production can be of great importance. In recent decades, protein engineering and genetic engineering have resulted in a high level of expression and production of this protein in a variety of hosts, including Escherichia coli bacteria using new techniques and methodes, hormone purification and assay are carried out easily. Therefore, the aim of this review was to investigate the production of recombinant human growth hormone (rhGH) and future challenges. One of the problems of the expression and purification of the human growth hormone may involve that maybe noted the production of inclusion bodies in the expression of recombinant proteins in the cell cytoplasm, the contamination caused by host proteins, low protein recovery from these inclusion bodies, low protein secretion into the Periplasmic space, high cost of production, especially in Purification stage and so on. Due to the lack of need for glycosylated hormone and high efficiency and simplicity of work, bacterial systems, especially Escherichia coli, are the most economical and effective systems for the expression of heterologous proteins. The hormone purification stage is usually the most costly process. Therefore, an optimal design for achieving the highest target protein recovery with the elimination of all contamination from the final product and reducing the purification step is required.

The aim of the present study was to map the morphological traits in Iranian Basil accessions (Oscillum Oscillos) by Inter Simple Sequence Repeats (ISSRs) markers.  In this experimental study, 50 Iranian basil accessions from different geographical regions were used and the experiment was based on a completely randomized design. Extracting DNA and PCR was performed with 12 ISSR primers for Basil accessions. Components of variance, general heritability, and genetic and phenotypic variation coefficients were calculated by formula. Bayesian method, linear mixed model as well as Minitab 17, DARwin 5, Structure 2.3.3, Tassel 3, and SPSS 20 software were used.  There was a positive correlation between the majority of traits for basil accessions. The highest coefficient of genetic variation was observed in stem diameter and internode length and the lowest was observed in flower number. Heritability varied between 3.63% and 94.24%. Foutrteen loci with 7 traits were recognized. The range of phenotypic changes varied from 3% to 29%. The highest number of locus was obtained for stem diameter traits and the lowest was obtained for lateral branch number. Six loci were specifically associated with only one trait and other loci were common in traits. The phenotypic variation varied between 17% and 29%.  Traits have a wide variety in Basil accessions and there is a positive correlation between the majority of them. The heritability of the traits varies from 3.63% to 94.24% and the range of the phenotypic changes varies from 3% to 29%. The highest number is for stem diameter traits and lowest is for lateral branch number. Six loci are specifically associated with only one trait and other loci are common in traits. The phenotypic variation varies between 29% ‐17%.

HIF-1 transcription factor is a key determinant of oxygen-dependent gene regulation, which its role has been demonstrated for the survival and progress of cancer tumors. The effect of suppression of HIF-1α on the evaluation of HIF-1 dependent processes and interference with pathophysiological events caused by hypoxia is important. The aim of this study was the apoptosis induction in glioma cells by downregulation of Hif-1α gene. In this experimental study, a specific siRNA against the HIF1α gene was developed using OligoWalk and Mit (siRNA.wi.mit.edu) servers and the online design department of Invivogene and Qiagene companies and the efficacy of its silencing in the U87 glioma cell line was quantitatively investigated by the Real-time PCR technique. In order to find out the effect of reduction of expression in the process of cell cycle and apoptosis, staining with PI and Annexin-PI was performed and the number of cells in each phase and the rate of cell mortality with control were compared by flow cytometry. The designed HIF-1a-siRNA was able to reduce HIF1α expression by 40%. The treatment of U87 cells after 24 hours increased the cells by 6% and after 48 hours, increased them by 12% in the sub G1 stage. Confirming the cell cycle changes, 48-hour treatment induced apoptosis in 58% of cells; regarding the 1.5% rate of apoptosis in the control cells, this cell death rate was very significant and showed the ability of the designed siRNA to induce apoptosis. The apoptosis induction of specific siRNA designed against HIF1α gene has a significant effect on the reduction of HIF-1α gene expression, cell growth, and apoptosis.

The aim of the present study was to isolate yeasts with the high ability of decolorization to use as biosorption in removing azo dyes. In this experimental study, an enrichment method was used to isolate dye absorbent yeast in a salt medium. The dye absorption was performed with comparing wet and dried biomass. Decolorization level was evaluated in different concentrations of dye and salt. By molecular method, the best strain was identified and its ability to absorb various dyes as well as mono-, di-, and tri-azo dyes were investigated. Statistical tests including one way ANOVA and Tukey as well as SPSS 19 software were used. Among 17 yeast isolates, ADH17 was selected as the most capable isolate. This isolate was 100% similar to Sarocladium sp. Dried biomass could adsorb the dye 4 times more than the wet biomass. The remained dye increased when initial dye concentration rose, but different concentrations of sodium chloride had no significant effect in biosorption. This strain could adsorb a broad range of azo dyes, including mono-, di-, and tri- azo and acidic, basic, and reactive dyes as well. The highest biosorption was 97.43% for reactive red and the lowest biosorption was 87.96% for reactive yellow. The ADH17 is the most capable isolate and it is 100% similar to Sarocladium sp. This strain adsorbs a broad range of azo dyes, including mono-, di-, and tri- azo and acidic, basic, and reactive dyes as well. Sarocladium sp has a high ability to absorb various azo dyes.

Biodiesel is considered as a clean fuel, because it is free of any aromatic compound. In recent years, in order to reduce the cost of production of Biodiesel, many studies have been conducted on the extraction of biofuels from microalgae around the world. Thus, this study was conducted with the aim of investigating the feasibility of optimum temperature for growth of Nannochloropsis Oculata microalga by using image processing system. In this experimental study, a piece of Nannochloropsis Oculata microalga containing 100,000 cells per ml was cultured in 15°C, 20°C, and 25°C. In order to evaluate the growth rate, active microalgae were sampled at 24-hour intervals, and their growth was studied, using machine vision systems. The data were analyzed, using Matlab 2012 and Weka 3 software by multivariable analysis of variance, linear regression algorithm, multilayer perceptron, Gaussian processing and simple linear regression analysis. The maximum cell density of Nannochloropsis Oculata on the 8th day was 286.23×104±0.38×105 cells per ml in treatment at 25°C and the minimum cell density was 168.58×104±0.48×105 cells per ml in treatment at 15°C. Specific growth rate was significantly increased at temperature of 25°C compared to the treatments at 15°C and 20°C. Linear regression algorithms (r2=0.84), multilayer perceptron (r2=0.88) and Gaussian processing (r2=0.78) showed good results, but simple linear regression indicated that the algorithm was unsuccessful (r2=0.45). The image processing technique provides a successful estimation of the growth process of Nannochloropsis Oculata at different temperature levels.

Proteinase K is an extracellular endopeptidase, which is secreted by Tritirachium album Limber and belongs to the serine endopeptidase class. This enzyme is extensively applied to protein-related studies. The present study aimed at evaluating the effect of urea, guanidine hydrochloride (GnHCl), and organic solvents on the kinetic activity of proteinase K enzyme. In this experimental study, kinetics studies were performed, using UV-Vis spectrophotometer on different concentrations of substrate, urea, and GnHCl at 40˚C and pH 7.4. Urea decreased the Vmax and Km of enzyme at 1 and 2molar concentrations, but at higher concentrations such as 3 and 4molar, it increased enzyme activity. GnHCl had an inhibitory effect on the enzyme activity, resulting in a decrease in Vmax and Km in 1, 2, and 3molar concentrations and acted as an uncompetitive inhibitor. Organic solvents including methanol, ethanol, and isopropanol had activatory effect at low concentrations and inhibitory effect at high concentrations on the kinetic activity of proteinase K enzyme. Urea has an inhibitory effect at low concentrations and an activatory effect on the activity of the enzyme at a concentrations above 2molar, but GnHCl has an inhibitory effect at all concentrations and can be used as an enzyme inhibitor. The effect of organic solvents including methanol, ethanol, and isopropanol on the activity of the proteinase K enzyme depends on their volume/volume percent; they cause enzyme activation at low percentages, but have inhibitory effect at high percentages, so that activates methanol below 30%  and isopropanol below 50%.

Mangroves are subjected to a range of abiotic stresses, which affect their growth and normal physiological processes. One of the most important modes of enzymatic antioxidant defense against stress caused by reactive oxygen species (ROS) is superoxide dismutase (SOD). The aim of this study was to evaluate the antioxidant enzymes activity of superoxide dismutase in the avicennia marina from the Persian Gulf and Gulf of Oman in the presence of the metal ions. In the present experimental study, which was conducted on the leaf of avicennia marina, the sampling was carried out from two habitats including Khamir port in the Persian Gulf and Sirik in the Gulf of Oman and the treatments were carried out in 3 replications. H2O2 sensitivity test and KCN test were used to determine the SOD type. The data were analyzed, using SPSS 19 software by multivariate analysis of variance and Duncan's multiple range test for comparing the means. The type of SOD enzyme was detected as Copper-zinc superoxide dismutase (Cu/Zn-SOD). There was no significant difference between different treatments of metals between two regions, and no interaction was observed between metal factor, concentration, and type of region. A strong inhibitory effect was observed in the presence of HgCl2 solution and a weak inhibitory effect was observed in the presence of ZnSo4, FeSo4, and MgCl2 solutions. Copper, manganese, and cobalt ions significantly increase the activity of the superoxide dismutase, while monovalent ions such as sodium and potassium have little effect on increasing SOD activity and the activity of the antioxidant enzymes of avicennia marina leaf from Khamir port in the Persian Gulf and Sirik in the Gulf of Oman is not different.

Microorganisms are present not only in common environment, but also in extreme environments. Salt lakes with near or at saturating salinity are spread all over the world. Urmia Salt Lake is one of these hypersaline environments. The present study aimed at evaluating prokaryotic diversity in hypersaline environment by culture-independent method. In this experimental study, different regions of Urmia Lake were sampled and the genomic material extracted from the water sample was used as a pattern for the amplification of 16S rDNA and a fragment of the bop gene via polymerase chain reaction. By cloning, each of the amplified fragments belonging to a single strain was amplified by T/A cloning vector. To further investigate the biodiversity of Haloarchaea, the biodiversity of bop gene was studied in addition to studying 16S rDNA. By cloning and sequencing, 6 bacteria genera, including Acaryochloris, Adhaeribacter, Brachybacterium, Gloeocapsopsis, Cesiribacter, and Bacillus were identified. Archaeal library belonged to 5 genera, including Halonotius, Halolamina, Haloquadratum, Halomicroarcula, and Halorhabdus. The clone libraries of bacterial belonged to 4 phyla, including Bacteroidetes, Cyanobacteria, Actinobacteria, and Firmicutes. . The clone libraries of bop gene (as a molecular marker) belonged to genera, including Halorubrum, Natrialba, Haloquadratum, and Natrinema. The bop phylogeny was closely related to the 16S rDNA phylogeny. By cloning and sequencing, 6 bacteria genera, including Acaryochloris, Adhaeribacter, Brachybacterium, Gloeocapsopsis, Cesiribacter, and Bacillus were identified. The bop phylogeny is closely related to the 16S rDNA phylogeny.

Identifying the structure and function of alpha-Synuclein protein can lead to the development of appropriate treatments for Parkinson disease. The aim of the current study was to investigate DNA cloning and the expression of alpha-Synuclein protein in E. coli. In this experimental study, the sequence of encoding alpha-Synuclein in pRK172 recombinant plasmid was amplified by Polymerase Chain Reaction (PCR), using best primers. The synthesized DNA was, then, digested by restriction enzymes and cloned into pET28a and recombinant plasmid was transferred into the expression strain of E. coli (BL21) by Calcium Chloride method. The expression of alpha-Synuclein gene was induced by Isopropyl-Beta-D-Thiogalactoside (IPTG) and the expression of alpha-Synuclein was investigated by SDS polyacrylamide gel electrophoresis (SDS-PAGE) method. Sequencing was done, using the ClustalW algorithm by the BioEdit 5.0.9 program. In products of DNA enzymatic digestive reactions and pET28a plasmid with restriction enzymes, the size of the fragments indicated the correctness of the enzymatic reactions. The synthesized DNA and pET28a plasmid were 407 and 5369 nucleotides, respectively. The translation of the sequence of the cloned fragment revealed a 100% similarity to the human alpha-Synuclein protein. In expressing the recombinant protein in comparison with negative control samples, adding IPTG increased the expression of alpha-Synuclein protein in all samples, especially 2 hours after induction. Most of alpha-Synuclein expressed from the pET28a-alpha-Synuclein plasmid accumulated in the bacteria as incorporated objects. The alpha-Synuclein protein is cloned into the pET28a plasmid and formation of the objects incorporated by alpha-Synuclein is confirmed by the expression of the pET28a-alpha-Synuclein system and paves the way for producing this protein in high scale.

Emerging sciences and technologies have huge potential in the field of innovation; therefore, they should be protected against large uncertainties caused by unknown. The aim of this study was to evaluate biotechnology forecasting innovation pathways based on its convergence with other technologies. In this systematic review, by the future-oriented assessment of biotechnology innovation pathways, future biotechnology strategies were developed at the national level. All potential applications of the future innovation pathways of this technology were identified in the combination and convergence with nanotechnologies, information, and cognitive science and technology. The strength and weakness of the effects and barriers in all areas of biotechnology were considered in terms of the short-, mid-, and long-term; in the same timeframe, the barriers to these technologies were identified in the field of combined dual technologies and ultimately for biotechnology itself, and future strategies for biotechnology were proposed based on 4 strategies, including ignorance, investment, exploitation, and opportunism. In the field of biotechnology- information technology- in the mid-term, the greatest impact was on improving the quality of human life, improving social outcomes, and increasing the level of innovation, and in the field of biotechnology- nanotechnology and biotechnology- cognitive science on improving the quality of human life, increasing security and defending power, and improving the positive social consequences. The highest number of applications is the mid-term. The "exploitation" strategy should be used in biotechnology- cognitive science and biotechnology- nanotechnology, respectively. The "investment" strategy should be the most widely used in the common areas of biology with information technology. In the common areas of biotechnology with nanotechnology and cognitive sciences, the most application is the “opportunism" strategy.