Search results for: protein
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 421

Search results for: protein

331 All Proteins Have a Basic Molecular Formula

Authors: Homa Torabizadeh

Abstract:

This study proposes a basic molecular formula for all proteins. A total of 10,739 proteins belonging to 9 different protein groups classified on the basis of their functions were selected randomly. They included enzymes, storage proteins, hormones, signalling proteins, structural proteins, transport proteins, immunoglobulins or antibodies, motor proteins and receptor proteins. After obtaining the protein molecular formula using the ProtParam tool, the H/C, N/C, O/C, and S/C ratios were determined for each randomly selected sample. In this case, H, N, O, and S coefficients were specified per carbon atom. Surprisingly, the results demonstrated that H, N, O, and S coefficients for all 10,739 proteins are similar and highly correlated. This study demonstrates that despite differences in the structure and function, all known proteins have a similar basic molecular formula CnH1.58 ± 0.015nN0.28 ± 0.005nO0.30 ± 0.007nS0.01 ± 0.002n. The total correlation between all coefficients was found to be 0.9999.

Keywords: Protein molecular formula, Basic unit formula, Protparam tool.

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330 Effect of Drought Stress and Selenium Spraying on Superoxide Dismotase Activity of Winter Rapeseed (Brassica napus L.) Cultivars

Authors: A.R. Pazoki, A. H. Shirani Rad, D. Habibi, F. Paknejad, S. Kobraee, N. Hadayat

Abstract:

In the other to Study of drought stress and Selenium spraying effect on superoxide dismotase (SOD) activity of rapeseed (Brassica napus L.) cultivars in Shahr-e-Rey region, an experiment carried out in Split factorial design in the basis of randomized complete blocks with 4 replications in 2006. Irrigation in two levels: Normal irrigation and irrigation with drought stress when the soil electrical conductivity reached to 60 as main factor and rapeseed cultivars in 3 levels Zarfam, Okapi, Opera and selenium spraying at the beginning of flowering stage in 3 levels: 0, 16 and 21 g/ha as sub factor. The results showed that the simple and interaction effect of irrigation, selenium and cultivars on SOD activity had significant difference. In this case Zarfam cultivar with 2010 u.mg-1 protein and Opera with 1454 u.mg-1 protein produced maximum and minimum amounts of SOD activitiy. Interaction effect of irrigation and variety showed that, normal irrigation in Opera with 1115 u.mg-1 protein and drought stress in Zarfam with 2784 u.mg-1 protein conducted to and minimum and maximum amounts of SOD activity. Interaction effect of irrigation, cultivar and selenium on SOD indicated that drought stress condition and 21 gr/ha selenium spraying in Zarfam variety with 3146 u.mg-1 protein gained to highest activities of SOD.

Keywords: Drought stress, Rapeseed, Selenium, Superoxide dismutase.

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329 Re-Engineering of Traditional Indian Wadi into Ready-to-Use High Protein Quality and Fibre Rich Chunk

Authors: Radhika Jain, Sangeeta Goomer

Abstract:

In the present study an attempt has been made to re-engineer traditional wadi into wholesome ready-to-use cereal-pulse-based chunks rich in protein quality and fibre content. Chunks were made using extrusion-dehydration combination. Two formulations i.e., whole green gram dhal with instant oats and washed green gram dhal with whole oats were formulated. These chunks are versatile in nature as they can be easily incorporated in day-to-day home-made preparations such as pulao, potato curry and kadhi. Cereal-pulse ratio was calculated using NDpCal%. Limiting amino acids such as lysine, tryptophan, methionine, cysteine and threonine were calculated for maximum amino acid profile in cereal-pulse combination. Time-temperature combination for extrusion at 130oC and dehydration at 65oC for 7 hours and 15 minutes were standardized to obtain maximum protein and fibre content. Proximate analysis such as moisture, fat and ash content were analyzed. Protein content of formulation was 62.10% and 68.50% respectively. Fibre content of formulations was 2.99% and 2.45%, respectively. Using a 5-point hedonic scale, consumer preference trials of 102 consumers were conducted and analyzed. Evaluation of chunks prepared in potato curry, kadi and pulao showed preferences for colour 82%, 87%, 86%, texture and consistency 80%, 81%, 88%, flavour and aroma 74%, 82%, 86%, after taste 70%, 75%, 86% and overall acceptability 77%, 75%, 88% respectively. High temperature inactivates antinutritional compounds such as trypsin inhibitors, lectins, saponins etc. Hence, availability of protein content was increased. Developed products were palatable and easy to prepare.

Keywords: Extrusion, NDpCal%, protein quality, wadi.

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328 An Evaluation of Pesticide Stress Induced Proteins in three Cyanobacterial Species-Anabaena Fertilissima, Aulosira Fertilissima and Westiellopsis Prolifica using SDS-PAGE

Authors: Nirmal Kumar, Rita N. Kumar, Anubhuti Bora, Manmeet Kaur Amb

Abstract:

The whole-cell protein-profiling technique was evaluated for studying differences in banding pattern of three different species of Cyanobacteria i.e. Anabaena fertilissima, Aulosira fertilissima and Westiellopsis prolifica under the influence of four different pesticides-2,4-D (Ethyl Ester of 2,4-Dichloro Phenoxy Acetic Acid), Pencycuron (N-[(4-chlorophenyl)methyl]-Ncyclopentyl- N'–phenylurea), Endosulfan (6,7,8,9,10,10hexachloro- 1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepine-3- oxide) and Tebuconazole (1-(4-Chlorophenyl)-4,4-dimethyl-3-(1,2,4- triazol-1-ylmethyl)pentan-3-ol). Whole-cell extracts were obtained by sonication treatment (Sonifier cell disruptor -Branson Digital Sonifier S-450D, USA) and were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGE analyses of the total protein profile of Anabaena fertilissima, Aulosira fertilissima and Westiellopsis prolifica showed a linear decrease in the protein content with increasing pesticide stress when administered to different concentrations of 2, 4-D, Pencycuron, Endosulfan and Tebuconazole. The results indicate that different stressors exert specific effects on cyanobacterial protein synthesis.

Keywords: Cyanobacteria, pesticide, SDS-PAGE

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327 Statistics of Exon Lengths in Animals, Plants, Fungi, and Protists

Authors: Alexander Kaplunovsky, Vladimir Khailenko, Alexander Bolshoy, Shara Atambayeva, AnatoliyIvashchenko

Abstract:

Eukaryotic protein-coding genes are interrupted by spliceosomal introns, which are removed from the RNA transcripts before translation into a protein. The exon-intron structures of different eukaryotic species are quite different from each other, and the evolution of such structures raises many questions. We try to address some of these questions using statistical analysis of whole genomes. We go through all the protein-coding genes in a genome and study correlations between the net length of all the exons in a gene, the number of the exons, and the average length of an exon. We also take average values of these features for each chromosome and study correlations between those averages on the chromosomal level. Our data show universal features of exon-intron structures common to animals, plants, and protists (specifically, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Cryptococcus neoformans, Homo sapiens, Mus musculus, Oryza sativa, and Plasmodium falciparum). We have verified linear correlation between the number of exons in a gene and the length of a protein coded by the gene, while the protein length increases in proportion to the number of exons. On the other hand, the average length of an exon always decreases with the number of exons. Finally, chromosome clustering based on average chromosome properties and parameters of linear regression between the number of exons in a gene and the net length of those exons demonstrates that these average chromosome properties are genome-specific features.

Keywords: Comparative genomics, exon-intron structure, eukaryotic clustering, linear regression.

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326 Differentiation between Common Tick Species Using Molecular Biology Techniques in Saudi Arabia

Authors: Kholoud A. Al-Shammery , Badr El-Sabah A. Fetoh, Ahmed M. Alshammari

Abstract:

Protein and Esterase electrophoresis were used to genetically identify two Saudi tick species. Engorged females of the camel tick Hyalomma dromedarii (Koch) (Acari: Ixodidae) and the cattle tick Boophilus annulatus (Say) (Acari: Ixodidae) ticks collected from infested camels and cattle in the animals resting house at Hail region in KSA were used. The results showed that there are a variation in both of protein and esterase activity levels and a high polymorphism within and between the genera and species of Hyalomma and Boophilus . In conclusion, the protein and esterase electrophoretic analysis used in the present study could successfully distinguish among tick species, commonly found in Saudi Arabia.

Keywords: Molecular biology, The camel tick Hyalomma dromedarii, The cattle tick Boophilus annulatus , Ticks.

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325 Production of H5N1 Hemagglutinin inTrichoplusia ni Larvae by a Novel Bi-cistronic Baculovirus Expression Vector

Authors: Tzyy Rong Jinn, Nguyen Tiep Khac, Tzong Yuan Wu

Abstract:

Highly pathogenic avian influenza (HPAI) H5N1 viruses have created demand for a cost-effective vaccine to prevent a pandemic of the disease. Here, we report that Trichoplusia ni (T. ni) larvae can act as a cost-effective bioreactor to produce recombinant HA5 (rH5HA) proteins as an potential effective vaccine for chickens. To facilitate the recombinant virus identification, virus titer determination and access the infected larvae, we employed the internal ribosome entry site (IRES) derived from Perina nuda virus (PnV, belongs to insect picorna like Iflavirus genus) to construct a bi-cistronic baculovirus expression vector that can express the rH5HA protein and enhanced green fluorescent protein (EGFP) simultaneously. Western blot analysis revealed that the 70 kDa rH5HA protein and partially cleaved products (40 kDa H5HA1) were generated in T. ni larvae infected with recombinant baculovirus carrying the H5HA gene. These data suggest that the baculovirus-larvae recombinant protein expression system could be a cost-effective platform for H5N1 vaccine production.

Keywords: Avian Influenza, baculovirus, hemagglutinin, Trichoplusia ni larvae

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324 Comparison of Nutritional and Chemical Parameters of Soymilk and Cow milk

Authors: Bahareh Hajirostamloo

Abstract:

Cow milk, is a product of the mammary gland and soymilk is a beverage made from soybeans; it is the liquid that remains after soybeans are soaked. In this research effort, we compared nutritional parameters of this two kind milk such as total fat, fiber, protein, minerals (Ca, Fe and P), fatty acids, carbohydrate, lactose, water, total solids, ash, pH, acidity and calories content in one cup (245 g). Results showed soymilk contains 4.67 grams of fat, 0.52 of fatty acids, 3.18 of fiber, 6.73 of protein, 4.43 of carbohydrate, 0.00 of lactose, 228.51 of water, 10.40 of total solids and 0.66 of ash, also 9.80 milligrams of Ca, 1.42 of Fe, and 120.05 of P, 79 Kcal of calories, pH=6.74 and acidity was 0.24%. Cow milk contains 8.15 grams of fat, 5.07 of fatty acids, 0.00 of fiber, 8.02 of protein, 11.37 of carbohydrate, ´Çá4.27 of lactose, 214.69 of water, 12.90 of total solids, 1.75 of ash, 290.36 milligrams of Ca, 0.12 of Fe, and 226.92 of P, 150 Kcal of calories, pH=6.90 and acidity was 0.21% . Soy milk is one of plant-based complete proteins and cow milk is a rich source of nutrients as well. Cow milk is containing near twice as much fat as and ten times more fatty acids do soymilk. Cow milk contains greater amounts of mineral (except Fe) it contain more than three hundred times the amount of Ca and nearly twice the amount of P as does soymilk but soymilk contains more Fe (ten time more) than does cow milk. Cow milk and soy milk contain nearly identical amounts of protein and water and fiber is a big plus, dairy has none. Although what we choose to drink is really a mater of personal preference and our health objectives but looking at the comparison, soy looks like healthier choices.

Keywords: Soymilk, cow milk, nutritional, comparison.

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323 Sequence-based Prediction of Gamma-turn Types using a Physicochemical Property-based Decision Tree Method

Authors: Chyn Liaw, Chun-Wei Tung, Shinn-Jang Ho, Shinn-Ying Ho

Abstract:

The γ-turns play important roles in protein folding and molecular recognition. The prediction and analysis of γ-turn types are important for both protein structure predictions and better understanding the characteristics of different γ-turn types. This study proposed a physicochemical property-based decision tree (PPDT) method to interpretably predict γ-turn types. In addition to the good prediction performance of PPDT, three simple and human interpretable IF-THEN rules are extracted from the decision tree constructed by PPDT. The identified informative physicochemical properties and concise rules provide a simple way for discriminating and understanding γ-turn types.

Keywords: Classification and regression tree (CART), γ-turn, Physicochemical properties, Protein secondary structure.

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322 A Novel Strategy for Oriented Protein Immobilization

Authors: Ching-Wei Tsai, Chih-I Liu, Ruoh-Chyu Ruaana

Abstract:

A new strategy for oriented immobilization of proteins was proposed. The strategy contains two steps. The first step is to search for a docking site away from the active site on the protein surface. The second step is trying to find a ligand that is able to grasp the targeted site of the protein. To avoid ligand binding to the active site of protein, the targeted docking site is selected to own opposite charges to those near the active site. To enhance the ligand-protein binding, both hydrophobic and electrostatic interactions need to be included. The targeted docking site should therefore contain hydrophobic amino acids. The ligand is then selected through the help of molecular docking simulations. The enzyme α-amylase derived from Aspergillus oryzae (TAKA) was taken as an example for oriented immobilization. The active site of TAKA is surrounded by negatively charged amino acids. All the possible hydrophobic sites on the surface of TAKA were evaluated by the free energy estimation through benzene docking. A hydrophobic site on the opposite side of TAKA-s active site was found to be positive in net charges. A possible ligand, 3,3-,4,4- – Biphenyltetra- carboxylic acid (BPTA), was found to catch TAKA by the designated docking site. Then, the BPTA molecules were grafted onto silica gels and measured the affinity of TAKA adsorption and the specific activity of thereby immobilized enzymes. It was found that TAKA had a dissociation constant as low as 7.0×10-6 M toward the ligand BPTA on silica gel. The increase in ionic strength has little effect on the adsorption of TAKA, which indicated the existence of hydrophobic interaction between ligands and proteins. The specific activity of the immobilized TAKA was compared with the randomly adsorbed TAKA on primary amine containing silica gel. It was found that the orderly immobilized TAKA owns a specific activity twice as high as the one randomly adsorbed by ionic interaction.

Keywords: Protein Oriented immobilization, Molecular docking, ligand design, surface modification.

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321 Nutritional Evaluation of Sorghum Flour (Sorghumbicolor L. Moench) During Processing of Injera

Authors: Noha A. Mohammed, Isam A. Mohamed Ahmed, Elfadil E. Babiker

Abstract:

The present study was carried out to evaluate the nutritional value of sorghum flour during processing of injera (unleavened thick bread). The proximate composition of sorghum flour before and after fermentation and that of injera was determined. Compared to the raw flour and fermented one, injera had low protein (11.55%), ash (1.57%) and fat (2.40%) contents but high in fiber content. Moreover, injera was found to have significantly (P ≤ 0.05) higher energy (389.08 Kcal/100g) compared to raw and fermented sorghum flour. Injera contained lower levels of anti-nutritional factors (polyphenols, phytate and tannins) compared to raw and fermented sorghum. Also it was found to be rich in Ca (4.75mg/100g), Fe (3.95 mg/100g), and Cu (0.7 mg/100g) compared to that of raw and fermented flour. Moreover, both the extractable minerals and protein digestibility were high for injera due to low amount of anti-nutrients. Injera was found to contain an appreciable amount of amino acids except arginine and tyrosine.

Keywords: Cooking, Fermentation, Malt, Protein fractions, Sorghum.

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320 Construction of Recombinant E.coli Expressing Fusion Protein to Produce 1,3-Propanediol

Authors: Rosarin Rujananon, Poonsuk Prasertsan, Amornrat Phongdara, Tanate Panrat, Jibin Sun, Sugima Rappert, An-Ping Zeng

Abstract:

In this study, a synthetic pathway was created by assembling genes from Clostridium butyricum and Escherichia coli in different combinations. Among the genes were dhaB1 and dhaB2 from C. butyricum VPI1718 coding for glycerol dehydratase (GDHt) and its activator (GDHtAc), respectively, involved in the conversion of glycerol to 3-hydroxypropionaldehyde (3-HPA). The yqhD gene from E.coli BL21 was also included which codes for an NADPHdependent 1,3-propanediol oxidoreductase isoenzyme (PDORI) reducing 3-HPA to 1,3-propanediol (1,3-PD). Molecular modeling analysis indicated that the conformation of fusion protein of YQHD and DHAB1 was favorable for direct molecular channeling of the intermediate 3-HPA. According to the simulation results, the yqhD and dhaB1 gene were assembled in the upstream of dhaB2 to express a fusion protein, yielding the recombinant strain E. coliBL21 (DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP41Y3). Strain BP41Y3 gave 10-fold higher 1,3-PD concentration than E. coliBL21 (DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP31Y2) expressing the recombinant enzymes simultaneously but in a non-fusion mode. This is the first report using a gene fusion approach to enhance the biological conversion of glycerol to the value added compound 1,3- PD.

Keywords: Recombinant E.coli, 1, 3-propanediol, glycerol, fusion protein.

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319 A Probability based Pair Extension Method in Protein 2-DE Gel Image Analysis

Authors: Yanhua Jin, Won Suk Lee

Abstract:

The two-dimensional gel electrophoresis method (2-DE) is widely used in Proteomics to separate thousands of proteins in a sample. By comparing the protein expression levels of proteins in a normal sample with those in a diseased one, it is possible to identify a meaningful set of marker proteins for the targeted disease. The major shortcomings of this approach involve inherent noises and irregular geometric distortions of spots observed in 2-DE images. Various experimental conditions can be the major causes of these problems. In the protein analysis of samples, these problems eventually lead to incorrect conclusions. In order to minimize the influence of these problems, this paper proposes a partition based pair extension method that performs spot-matching on a set of gel images multiple times and segregates more reliable mapping results which can improve the accuracy of gel image analysis. The improved accuracy of the proposed method is analyzed through various experiments on real 2-DE images of human liver tissues.

Keywords: Proteomics, spot-matching, two-dimensionalelectrophoresis.

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318 Vetiver Oil Production from Root Culture of Vetiveria zizanioides

Authors: Rizkita R. Esyanti, Iriawati, Olga Mardisadora

Abstract:

Vetiver oil is secondary metabolite that accumulates in Vetiveria zizanioides roots.  The aim of this study was to obtain best type of root culture which produce high amount of vetiver oil, and was similar to metabolites produce from its mother plant.  Protein analysis was also conducted to detect protein, related to putative enzymes, which have a role in terpenoids synthesis in the root culture. The results showed that root culture derived from crown explant produced the best root growth.   The root culture produced primary and lateral roots, ca. 40 branches.  The vetiver oil produced from root culture was analyzed by using GC-MS., and the highest content of terpenoids from roots of crown explant attained 19.024%.  The result of SDS PAGE showed proteins which were ±61 kD and ± 68 kD, each might be related to putative monoterpene synthase and sesquiterpenes complex, respectively. 

Keywords: Protein, Root culture, Terpenoids, Vetiver oil.

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317 Virulent-GO: Prediction of Virulent Proteins in Bacterial Pathogens Utilizing Gene Ontology Terms

Authors: Chia-Ta Tsai, Wen-Lin Huang, Shinn-Jang Ho, Li-Sun Shu, Shinn-Ying Ho

Abstract:

Prediction of bacterial virulent protein sequences can give assistance to identification and characterization of novel virulence-associated factors and discover drug/vaccine targets against proteins indispensable to pathogenicity. Gene Ontology (GO) annotation which describes functions of genes and gene products as a controlled vocabulary of terms has been shown effectively for a variety of tasks such as gene expression study, GO annotation prediction, protein subcellular localization, etc. In this study, we propose a sequence-based method Virulent-GO by mining informative GO terms as features for predicting bacterial virulent proteins. Each protein in the datasets used by the existing method VirulentPred is annotated by using BLAST to obtain its homologies with known accession numbers for retrieving GO terms. After investigating various popular classifiers using the same five-fold cross-validation scheme, Virulent-GO using the single kind of GO term features with an accuracy of 82.5% is slightly better than VirulentPred with 81.8% using five kinds of sequence-based features. For the evaluation of independent test, Virulent-GO also yields better results (82.0%) than VirulentPred (80.7%). When evaluating single kind of feature with SVM, the GO term feature performs much well, compared with each of the five kinds of features.

Keywords: Bacterial virulence factors, GO terms, prediction, protein sequence.

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316 Investigation of Possible Behavioural and Molecular Effects of Mobile Phone Exposure on Rats

Authors: Ç. Gökçek-Saraç, Ş. Özen, N. Derin

Abstract:

The N-methyl-D-aspartate (NMDA)-dependent pathway is the major intracellular signaling pathway implemented in both short- and long-term memory formation in the hippocampus which is the most studied brain structure because of its well documented role in learning and memory. However, little is known about the effects of RF-EMR exposure on NMDA receptor signaling pathway including activation of protein kinases, notably Ca2+/calmodulin-dependent protein kinase II alpha (CaMKIIα). The aim of the present study was to investigate the effects of acute and chronic 900 MHz RF-EMR exposure on both passive avoidance behaviour and hippocampal levels of CaMKIIα and its phosphorylated form (pCaMKIIα). Rats were divided into the following groups: Sham rats, and rats exposed to 900 MHz RF-EMR for 2 h/day for 1 week (acute group) or 10 weeks (chronic group), respectively. Passive avoidance task was used as a behavioural method. The hippocampal levels of selected kinases were measured using Western Blotting technique. The results of passive avoidance task showed that both acute and chronic exposure to 900 MHz RF-EMR can impair passive avoidance behaviour with minor effects on chronic group of rats. The analysis of western blot data of selected protein kinases demonstrated that hippocampal levels of CaMKIIα and pCaMKIIα were significantly higher in chronic group of rats as compared to acute groups. Taken together, these findings demonstrated that different duration times (1 week vs 10 weeks) of 900 MHz RF-EMR exposure have different effects on both passive avoidance behaviour of rats and hippocampal levels of selected protein kinases.

Keywords: Hippocampus, protein kinase, rat, RF-EMR.

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315 Protein Production by Bacillus Subtilis Atcc 21332 in the Presence of Cymbopogon Essential Oils

Authors: Hanina M. N., Hairul Shahril M., Mohd Fazrullah Innsan M. F., Ismatul Nurul Asyikin I., Abdul Jalil A. K, Salina M. R., Ahmad I.B.

Abstract:

Proteins levels produced by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antimicrobial agents or antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics or natural compounds in nature as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was focusing on the effect of essential oils from Cymbopogon flexuosus and C. nardus in regulating proteins production by Bacillus subtilis ATCC 21332. The Minimum Inhibition Concentrations (MICs) of both essential oils on B. subtilis were determined by using microdilution assay, resulting 0.2% and 1.56% for each C. flexuosus and C. nardus subsequently. The bacteria were further exposed to each essential oils at concentration of 0.01XMIC for 2 days. The proteins were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Protein profile showed that a band with approximate size of 250 kD was appeared for the treated bacteria with essential oils. Thus, Bacillus subtilis ATCC 21332 in stressful condition with the presence of essential oils at low concentration could induce the protein production.

Keywords: Bacillus subtilis ATCC 21332, Cymbopogon essential oils, protein

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314 The Role of Immunogenic Adhesin Vibrio alginolyticus 49 k Da to Molecule Expression of Major Histocompatibility Complex on Receptors of Humpback Grouper Cromileptes altivelis

Authors: Uun Yanuhar

Abstract:

The purpose of research was to know the role of immunogenic protein of 49 kDa from V.alginolyticus which capable to initiate molecule expression of MHC Class II in receptor of Cromileptes altivelis. The method used was in vivo experimental research through testing of immunogenic protein 49 kDa from V.alginolyticus at Cromileptes altivelis (size of 250 - 300 grams) using 3 times booster by injecting an immunogenic protein in a intramuscular manner. Response of expressed MHC molecule was shown using immunocytochemistry method and SEM. Results indicated that adhesin V.alginolyticus 49 kDa which have immunogenic character could trigger expression of MHC class II on receptor of grouper and has been proven by staining using immunocytochemistry and SEM with labeling using antibody anti MHC (anti mouse). This visible expression based on binding between epitopes antigen and antibody anti MHC in the receptor. Using immunocytochemistry, intracellular response of MHC to in vivo induction of immunogenic adhesin from V.alginolyticus was shown.

Keywords: C.altivelis, immunogenic, MHC, V.alginolyticus.

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313 Proteolytic Dedradation of Anchovy (Spolephorus spp.) Proteins by Halophilic Proteinase from Halobacillus sp. SR5-3

Authors: Sirilak Namwong , Wonnop Visessanguan, Soottawat Benjakul, Somboon Tanasupawat

Abstract:

The halophilic proteinase showed a maximal activity at 50°C and pH 9~10, in 20% NaCl and was highly stabilized by NaCl. It was able to hydrolyse natural actomyosin (NAM), collagen and anchovy protein. For NAM hydrolysis, the myosin heavy chain was completely digested by halophilic proteinase as evidenced by the lowest band intensity remaining, but partially hydrolysed actin. The SR5-3 proteinase was also capable hydrolyzing two major components of collagen, β- and α-compounds, effectively. The degree of hydrolysis (DH) of the halophilic proteinase and commercial proteinases (Novozyme, Neutrase, chymotrypsin and Flavourzyme) on the anchovy protein, were compared, and it was found that the proteinase showed a greater degree of hydrolysis towards anchovy protein than that from commercial proteinases. DH of halophilic proteinase was sharply enhanced according to the increase in the concentration of enzyme from 0.035 U to 0.105 U. The results warranting that the acceleration of the production of fish sauce with higher quality, may be achieved by adding of the halophilic proteinase from this bacterium.

Keywords: Halophilic proteinase, Halobacillus sp. SR5-3, anchovy (Spolephorus spp.) proteins, fish sauce

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312 Cloning, Expression and Protein Purification of AV1 Gene of Okra Leaf Curl Virus Egyptian Isolate and Genetic Diversity between Whitefly and Different Plant Hosts

Authors: Dalia. G. Aseel

Abstract:

Begomoviruses are economically important plant viruses that infect dicotyledonous plants and exclusively transmitted by the whitefly Bemisia tabaci. Here, replicative form was isolated from Okra, Cotton, Tomato plants and whitefly infected with Begomoviruses. Using coat protein specific primers (AV1), the viral infection was verified with amplicon at 450 bp. The sequence of OLCuV-AV1 gene was recorded and received an accession number (FJ441605) from Genebank. The phylogenetic tree of OLCuV was closely related to Okra leaf curl virus previously isolated from Cameroon and USA with nucleotide sequence identity of 92%. The protein purification was carried out using His-Tag methodology by using Affinity Chromatography. The purified protein was separated on SDS-PAGE analysis and an enriched expected size of band at 30 kDa was observed. Furthermore, RAPD and SDS-PAGE were used to detect genetic variability between different hosts of okra leaf curl virus (OLCuV), cotton leaf curl virus (CLCuV), tomato yellow leaf curl virus (TYLCuV) and the whitefly vector. Finally, the present study would help to understand the relationship between the whitefly and different economical crops in Egypt.

Keywords: Begomovirus, AV1 gene, sequence, cloning, whitefly, okra, cotton, tomato, RAPD, phylogenetic tree and SDS-PAGE.

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311 Proximate and Mineral Composition of Chicken Giblets from Vojvodina (Northern Serbia)

Authors: M. R. Jokanović, V. M. Tomović, M. T. Jović, S. B. Škaljac, B. V. Šojić, P. M. Ikonić, T. A. Tasić

Abstract:

Proximate (moisture, protein, total fat, total ash) and mineral (K, P, Na, Mg, Ca, Zn, Fe, Cu and Mn) composition of chicken giblets (heart, liver and gizzard) were investigated. Phosphorous content, as well as proximate composition, were determined according to recommended ISO methods. The content of all elements, except phosphorus, of the giblets tissues were determined using inductively coupled plasma-optical emission spectrometry (ICP-OES), after dry ashing mineralization. Regarding proximate composition heart was the highest in total fat content, and the lowest in protein content. Liver was the highest in protein and total ash content, while gizzard was the highest in moisture and the lowest in total fat content. Regarding mineral composition liver was the highest for K, P, Ca, Mg, Fe, Zn, Cu, and Mn, while heart was the highest for Na content. The contents of almost all investigated minerals in analysed giblets tissues of chickens from Vojvodina were similar to values reported in the literature, i.e. in national food composition databases of other countries.

Keywords: Chicken giblets, proximate composition, mineral composition.

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310 More Realistic Model for Simulating Min Protein Dynamics: Lattice Boltzmann Method Incorporating the Role of Nucleoids

Authors: J.Yojina, W. Ngamsaad, N. Nuttavut, D.Triampo, Y. Lenbury, W. Triampo, P. Kanthang, S.Sriyab

Abstract:

The dynamics of Min proteins plays a center role in accurate cell division. Although the nucleoids may presumably play an important role in prokaryotic cell division, there is a lack of models to account for its participation. In this work, we apply the lattice Boltzmann method to investigate protein oscillation based on a mesoscopic model that takes into account the nucleoid-s role. We found that our numerical results are in reasonably good agreement with the previous experimental results On comparing with the other computational models without the presence of nucleoids, the highlight of our finding is that the local densities of MinD and MinE on the cytoplasmic membrane increases, especially along the cell width, when the size of the obstacle increases, leading to a more distinct cap-like structure at the poles. This feature indicated the realistic pattern and reflected the combination of Min protein dynamics and nucleoid-s role.

Keywords: lattice Boltzmann method, cell division, Minproteins oscillation, nucleoid

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309 Mutational Analysis of CTLA4 Gene in Pakistani SLE Patients

Authors: N. Hussain, G. Jaffery, A.N. Sabri, S. Hasnain

Abstract:

The main aim is to perform mutational analysis of CTLA4 gene Exon 1 in SLE patients. A total of 61 SLE patients fulfilling “American College of Rheumatology (ACR) criteria" and 61 controls were enrolled in this study. The region of CTLA4 gene exon 1 was amplified by using Step-down PCR technique. Extracted DNA of band 354 bp was sequenced to analyze mutations in the exon-1 of CTLA-4 gene. Further, protein sequences were identified from nucleotide sequences of CTLA4 Exon 1 by using Expasy software and through Blast P software it was found that CTLA4 protein sequences of Pakistani SLE patients were similar to that of Chinese SLE population. No variations were found after patients sequences were compared with that of the control sequence. Furthermore it was found that CTLA4 protein sequences of Pakistani SLE patients were similar to that of Chinese SLE population. Thus CTLA4 gene may not be responsible for an autoimmune disease SLE.

Keywords: American College of Rheumatology criteria, autoimmune disease, Cytotoxic T Lymphocyte Antigen-4, Polymerase Chain Reaction, Systemic Lupus Erythematosus

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308 Computational Analysis of Potential Inhibitors Selected Based On Structural Similarity for the Src SH2 Domain

Authors: W. P. Hu, J. V. Kumar, Jeffrey J. P. Tsai

Abstract:

The inhibition of SH2 domain regulated protein-protein interactions is an attractive target for developing an effective chemotherapeutic approach in the treatment of disease. Molecular simulation is a useful tool for developing new drugs and for studying molecular recognition. In this study, we searched potential drug compounds for the inhibition of SH2 domain by performing structural similarity search in PubChem Compound Database. A total of 37 compounds were screened from the database, and then we used the LibDock docking program to evaluate the inhibition effect. The best three compounds (AP22408, CID 71463546 and CID 9917321) were chosen for MD simulations after the LibDock docking. Our results show that the compound CID 9917321 can produce a more stable protein-ligand complex compared to other two currently known inhibitors of Src SH2 domain. The compound CID 9917321 may be useful for the inhibition of SH2 domain based on these computational results. Subsequently experiments are needed to verify the effect of compound CID 9917321 on the SH2 domain in the future studies.

Keywords: Nonpeptide inhibitor, Src SH2 domain, LibDock, molecular dynamics simulation.

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307 The Expression of a Novel Gene Encoding an Ankyrin-Repeat Protein, DRA1, is Regulated by Drought-Responsive Alternative Splicing

Authors: H. Sakamoto, Y. Nakagawara, S. Oguri

Abstract:

Drought stress is a critical environmental factor that adversely affects crop productivity and quality. Because of their immobile nature, plants have evolved mechanisms to sense and respond to drought stress. We identified a novel locus of Arabidopsis, designated DRA1 (drought responsive ankyrin1), whose disruption leads to increased drought-stress tolerance. DRA1 encodes a transmembrane protein with an ankyrin-repeat motif that has been implicated in diverse cellular processes such as signal transduction. RT-PCR analysis revealed that there were at least two splicing variants of DRA1 transcripts in wild-type plants. In response to drought stress, the levels of DRA1 transcripts retaining second and third introns were increased, whereas these introns were removed under unstressed conditions. These results suggest that DRA1 protein may negatively regulate plant drought tolerance and that the expression of DRA1is regulated in response to drought stress by alternative splicing.

Keywords: Alternative splicing, ankyrin repeat, Arabidopsis, drought tolerance.

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306 The Expression of a Novel Gene Encoding an Ankyrin-Repeat Protein, DRA1, is Regulated by Drought-Responsive Alternative Splicing

Authors: H. Sakamoto, Y. Nakagawara, S. Oguri

Abstract:

Drought stress is a critical environmental factor that adversely affects crop productivity and quality. Because of their immobile nature, plants have evolved mechanisms to sense and respond to drought stress. We identified a novel locus of Arabidopsis, designated DRA1 (drought responsive ankyrin1), whose disruption leads to increased drought-stress tolerance. DRA1 encodes a transmembrane protein with an ankyrin-repeat motif that has been implicated in diverse cellular processes such as signal transduction. RT-PCR analysis revealed that there were at least two splicing variants of DRA1 transcripts in wild-type plants. In response to drought stress, the levels of DRA1 transcripts retaining second and third introns were increased, whereas these introns were removed under unstressed conditions. These results suggest that DRA1 protein may negatively regulate plant drought tolerance and that the expression of DRA1is regulated in response to drought stress by alternative splicing.

Keywords: Alternative splicing, ankyrin repeat, Arabidopsis, drought tolerance.

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305 Multi-Label Hierarchical Classification for Protein Function Prediction

Authors: Helyane B. Borges, Julio Cesar Nievola

Abstract:

Hierarchical classification is a problem with applications in many areas as protein function prediction where the dates are hierarchically structured. Therefore, it is necessary the development of algorithms able to induce hierarchical classification models. This paper presents experimenters using the algorithm for hierarchical classification called Multi-label Hierarchical Classification using a Competitive Neural Network (MHC-CNN). It was tested in ten datasets the Gene Ontology (GO) Cellular Component Domain. The results are compared with the Clus-HMC and Clus-HSC using the hF-Measure.

Keywords: Hierarchical Classification, Competitive Neural Network, Global Classifier.

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304 Expression of Tissue Plasminogen Activator in Transgenic Tobacco Plants by Signal Peptides Targeting for Delivery to Apoplast, Endoplasmic Reticulum and Cytosol Spaces

Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi

Abstract:

Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA4404. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.

Keywords: Recombinant tissue plasminogen activator, plant cell comportment, leader signals, transgenic tobacco.

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303 Alignment of MG-63 Osteoblasts on Fibronectin-Coated Phosphorous Doping Lattices in Silicon

Authors: Andreas Körtge, Susanne Stählke, Regina Lange, Mario Birkholz, Mirko Fraschke, Katrin Schulz, Barbara Nebe, Patrick Elter

Abstract:

A major challenge in biomaterials research is the regulation of protein adsorption which is a key factor for controlling the subsequent cell adhesion at implant surfaces. The aim of the present study was to control the adsorption of fibronectin (FN) and the attachment of MG-63 osteoblasts with an electronic nanostructure. Shallow doping line lattices with a period of 260 nm were produced for this purpose by implantation of phosphorous in silicon wafers. Protein coverage was determined after incubating the substrate with FN by means of an immunostaining procedure and the measurement of the fluorescence intensity with a TECAN analyzer. We observed an increased amount of adsorbed FN on the nanostructure compared to control substrates. MG-63 osteoblasts were cultivated for 24h on FN-incubated substrates and their morphology was assessed by SEM. Preferred orientation and elongation of the cells in direction of the doping lattice lines was observed on FN-coated nanostructures.

Keywords: Cell adhesion, electronic nanostructures, doping lattice, fibronectin, MG-63 osteoblasts, protein adsorption.

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302 Diabetes Mellitus and Food Balance in the Kingdom of Saudi Arabia

Authors: Aljabryn Dalal Hamad

Abstract:

The present explanatory study concerns with the relation between Diabetes Mellitus and Food Balance in the Kingdom of Saudi Arabia during 2005-2010, using published data. Results illustrated that Saudi citizen daily protein consumption (DPC) during 2005-2007 (g/capita/day) is higher than the average global consumption level of protein with 15.27%, daily fat consumption (DFC) with 24.56% and daily energy consumption (DEC) with 16.93% and increases than recommended level by International Nutrition Organizations (INO) with 56% for protein, 60.49% for fat and 27.37% for energy. On the other hand, DPC per capita in Saudi Arabia decreased during the period 2008-2010 from 88.3 to 82.36 gram/ day. Moreover, DFC per capita in Saudi Arabia decreased during the period 2008-2010 from 3247.90 to 3176.43 Cal/capita/ day, and daily energy consumption (DEC) of Saudi citizen increases than world consumption with 16.93%, whereas increases with 27.37% than INO. Despite this, DPC, DFC and DEC per capita in Saudi Arabia still higher than world mean. On the other side, results illustrated that the number of diabetic patients in Saudi Arabia during the same period (2005-2010). The curve of diabetic patient’s number in Saudi Arabia during 2005-2010 is regular ascending with increasing level ranged between 7.10% in 2005 and 12.44% in 2010. It is essential to devise Saudi National programs to educate the public about the relation of food balances and diabetes so it could be avoided, and provide citizens with healthy dietary balances tables.

Keywords: Diabetes Mellitus, Food Balance, Energy, Fat, Protein, Saudi Arabia.

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