Research Journal of Biotechnology

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Research Journal of Biotechnology





Isolation and Expression of Caspase-3 Genes in Different Development Stages, Tissues and Treatments of the Dastarcus helophoroides (Fairmaire) (Coleoptera: Bothrideridae)

Thiquynhtrang Vu, Thanh Pham and Menglou Li

The caspase is family of cysteine proteases that plays a central role in the initiation and execution phases of apoptosis. We isolated Dh-caspase-3 gene from Dastarcus helophoroides and found that it is highly homologous to effector caspase genes in other insect and mammalian species. Dh-caspase-3 had a full length of 1128 bp and contained 1008 bp open reading frame that encoded 335 amino acids. Dh-caspase-3 mRNA expression was elevated with development stage, tissues and high temperature treatment. Differential expression of Dh-caspase-3 showed significant differences among development stage and tissue distribution. Dh-caspase-3 mRNA transcription levels were over-expression in both three development stages (adult, pupae, larvae) after high temperature treatment. Based on the results of this study, the increases of Dh-caspase-3 mRNA transcription could be induced under high temperature conditions. So, we suggest that Dh-caspase-3 gene may play an important defensive role in the apoptosis process of D. helophoroides.

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Effect of chemical mutagens on character association in sesame (Sesamum indicum L.)

Sanghani J. M., Vadher K. J., Sanghani A. O., Ramani H. R., Raval S. S. and Sorathiya J. S.

An investigation was carried out to study the nature and magnitude of induced genetic variability in sesame (Sesamum indicum L.). Seeds of three sesame genotypes, GT-2, PARAM and VIKRANT were treated with four concentrations of ethyl methane sulphonate (0.5%, 1.0%, 1.5% and 2.0%), separately. Mutant generations from M1 to M2 were raised to study the extent of variability, heritability and genetic advance in mutant populations. Mutations surpassed the magnitude of variability over control population in both the generations. Genotypic and phenotypic variances were higher for sterility % and 1000-seed weight in both M1 and M2 generations. Regardless of the genotype, M1 generation professed maximum genotypic and phenotypic coefficients of variability (GCV and PCV) for number of seeds per capsule. On the contrary, in M2 generation induced populations of all the three genotypes engendered maximum GCV and PCV for seed yield per plant. High heritability for sterility percentage and 1000-seed weight coupled with high genetic advance inferred that additive gene effects were important in determining these characters and could be improved through mass selection.

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Evaluation of genetic diversity in safflower (Carthamus tinctorius L.) using agro-morphological, fatty acid composition and ISSR molecular markers

Reza Talebi and Somayeh Ahangarian Abhari

Genetic diversity among 25 diverse genotypes of safflower (Carthamus tinctorius L.) was assumed using agro-morphological, fatty acid and ISSR markers. The results of analysis of variance analysis showed significant differences among accessions for all the agro-morphological and biochemical traits. Biological yield were significantly correlated with seed yields (0.83**) and had significant negative correlation with harvest index (-0.57**). Under the experimental conditions, oleic and linoleic acids together accounted for nearly 90% while stearic and palmitic acids together represented 9.5% of the total fatty acid concentration. The fatty acid somposition comprised 73.12% to 78.45% linoleic acid (C18:2), oleic acid 12.01% to 16.89%, 2.14% to 3.68% stearic acid (C18:0) and 0.27-0.55% linolenic acid (C18:3). Cluster analysis based on agro-morphological, biochemical and ISSR markers distinguished the genotypes into two, five and three distinct clusters respectively. High level of genetic diversity in this study indicates a considerable potential for improving safflower for both agronomic and quality traits.

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Cloning and expression of thioglucoside glucohydrolase MYR1 from tumorous stem mustard and optimization of enzymatic conditions

Yan Zhang and Yao Li

Myrosinase hydrolyzes glucosinolates to generate isothiocyanates (ITCs) that have potential as anti-cancer drugs and plant insecticides. To facilitate production of recombinant myrosinase, MYR1 from tumorous stem mustard was amplified by RT-PCR, cloned into the pPIC9K-S expression vector and recombinantly expressed in Pichia pastoris. Mycelia were cultured in YPD medium and expression induced by replacing with BMMY medium for 72 hours. SDS-PAGE confirmed that the 90 kDa myrosinase was purified by immobilized metal affinity chromatography to 85% purity with an overall yield of 850 U/l. With sinigrin as substrate, the specific activity was 0.003 U/μl. Using tumorous stem mustard isothiocyanates as standards, enzymatic conditions were investigated and 55C, pH 6.5, reaction time = 1 h, ascorbic acid content = 0.006 mg/g were optimal, yielding 1.665 mg/g isothiocyanates from glucosinolate substrates. Under identical conditions, 30% pure glucosinolate substrate mix generated the highest isothiocyanate content.

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Gene expression data classification using MapReduce version of KNN hybridized with PSO

Bhavani R. and Sudha Sadasivam G.

One of the important applications of gene expression data obtained from DNA microaaray technology is categorizing genes based on functionalities like disease type. This paper presents the classification of gene expression data using MapReduce version of k-nearest neighbors (KNN) hybridized with Particle Swarm Optimization (PSO). The top k-nearest neighbor genes are taken as input to the PSO algorithm to find the class labels. MapReduce programming is used to reduce the execution time for processing voluminous gene expression data. Experimental results show that the proposed method of gene expression data classification exhibits good scalability and improves classification accuracy by 5% to 7%.

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Evaluation of safety, antimicrobial activity and probiotic properties of Escherichia coli Nissle 1917 isolated from Idli batter

Chelliah Ramachandran, Ramakrishnan Sudha Rani and Antony Usha

The present study was undertaken to characterize probiotic efficiency and evaluate the safety of lactose utilizing Escherichia coli Nissle 1917 isolated from frozen idli batter. The strain inhibited the growth of 13 dysentery causing pathogens and shows commensal relationship with 6 lactic acid bacteria (LAB) strains, 4 commercial yeast and bacterial probiotic strains. PCR amplification with gram-negative 16S rRNA primers confirmed Escherichia coli Nissle 1917. The strain was resistant to physiological concentrations of bile salts, pepsin and pancreatic enzyme. It showed efficient auto-aggregation and co-aggregation ability with 6 LAB and 4 commercial probiotic yeast as well as exhibited good hydrophobicity in xylene and toluene. Isolated E. Nissle 1917 (KT000039) has probiotic characteristics similar to LAB and this may be a solution for the steadily increasing demand in food industry which has to overcome the need of efficient probiotics with a wide antimicrobial spectrum.

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Chondroitin sulfate modified Zirconium Phosphate nanoparticles for lung cancer chemotherapy

Wu Chungang, Long Shengyong and Liu Yougui

Novel tumor-targeting zirconium phosphate (ZP) nanoparticles modified with chondroitin sulfate (CS) were developed to explore the feasibility of exploiting the drug-loading property of ZP and the tumor-targeting ability of CS to construct a tumor-targeting cisplatin (CDDP) delivery system (CS-ZP) for potential lung cancer therapy. The experimental results indicated that CDDP loading into the CS-ZP nanoparticles was as high as 24.36% ± 5.37% which is beneficial to cancer therapy. CDDP-loaded CS-ZP nanoparticles increased the accumulation of CDDP in A549 lung cancer cells possibly via CD44 receptor-mediated endocytosis, which in turn, exhibited superior anti-cancer activity in vitro. In vivo anti-cancer efficacy assay also confirmed that CS-ZP nanoparticles possessed preferable anti-cancer ability which exhibited minimized toxic side effects of CDDP together with strong tumor-suppression potential in clinical application.

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Effect of Thermotherapy on Regeneration Response and Production of PLRV-Free Plants of Potato from Infected Tubers

Singh Balwinder

Potato leaf roll virus reduces crop yields and causes degeneration of seed stocks in potato growing regions of the world. Vegetative propagation of potato using virus infected seed tubers results into continuity and spread of this virus in next season crop. The present study was conducted to investigate efficacy of thermotherapy for elimination of PLRV from infected tubers along with evaluation of its effects on survival of tubers and regeneration response of sprouts. Thermotherapy was applied on virus infected tubers at 37, 38, 39, 40 and 41oC for 28 days. Sprouts obtained from heat treated tubers were given hot water treatment at same temperatures in water bath for 60, 90, 120, 150 and 180 min. Infected tubers, plants developed after thermotherapy of tubers and sprouts were indexed for the presence/absence of virus by DAS-ELISA and RT-PCR. Results showed that increase in temperature from 38 to 41oC during thermotherapy significantly decreases survival of tubers and regeneration response of sprouts. 54.85% tubers survived after treated at 41oC for 28 days and 16.45% of the total plants produced from these tubers were tested negative for PLRV in RT-PCR. Sprouts treated at 40oC for 180 min showed regeneration response of 31.94% and gave 39.58% PLRV free plants. Thermotherapy of sprouts at 40oC for 150 min improved regeneration response to 43.05% and 35.48% of the total plantlets obtained from this treatment were tested negative for PLRV. Heat treatment of tubers at 40oC for 28 days followed by sprouting and hot water treatment of sprouts at 40oC for 150 or 180 min was observed to be effective for production of PLRV free plants. Only those plants were considered as viruses free which were tested negative in both DAS-ELISA and RT-PCR.

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Cognitive Enhancing Activities of Marine Spirulina maxima from Ultrasonification Extraction Process

Lee Hyeon Yong

This study evaluated the cognitive-enhancing activities of the Spirulina maxima 70% ethanol extract through ultrasonification extraction process at 400C (UE)compared to Chlorophyll a with Morris water maze and passive avoidance tests in mice. The cytotoxicity of the S. maxima extract obtained by ultrasonification extraction showed low cytotoxicity as 15.70% against murine HT22 cells. In particular, the highest anti-oxidant activities of DPPH were measured as 35.35% compared with 30.00% in the extract from conventional ethanol extraction at 800C (EE). It was clearly shown that the UE enhanced cognitive function, compared to the EE in feeding both dosages of 200 and 400 mg/kg of mice. Specifically, the extract from ultrasonification process was shown by a decrease in escape latency time and increased latency time. Interestingly, Chlorophyll a was also associated with good cognitive activities in the Morris water maze and the passive avoidance test in mice treated at10 mg/kg. In this work, it was reported that the UE improved cognitive functions and was also superior to the EE, possibly due to less destruction and denaturation of biologically active substances by being processed at low temperature. It was also proved that activities of the UE could be comparable to Chlorophyll a that is known to have cognitive activity.

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Lipid Peroxidation and Biochemical Abnormalities in Tannery Workers exposed to Hexavalent Chromium

Ateeq Muhammad and Ali Fawad

Chromate salts are extensively used in tanning industries; the oxidation process, high temperature and pH in tanning environment converts the Cr (III) into chromium (VI) which is more carcinogenetic and toxic for genetic material in cell free system. The aim of the present study was to investigate whether exposure to hexavalent chromium induces lipid peroxidation and biochemical abnormalities in tannery workers. Chromium level in erythrocytes was determined by by graphite furnace atomic absorption spectrophotometer. The results showed that blood chromium level, superoxide dismutase (SOD) and malondialdehyde (MDA) levels were significantly higher (p < 0.001) while the level of glutathione (GSH) was significantly lower (p < 0.001) in exposed groups compared with control group. The values of liver function tests of tannery workers were found to be within the normal range in all age groups except for the albumin content and alkaline phosphatase (AP) activity. AP activity was significantly higher in both the exposed groups I and II. Albumin level was lowered in both the exposed groups I and II. The present study revealed that prolonged exposure to hexavalent chromium is likely to induce lipid peroxidation and biochemical abnormalities in tannery workers.

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Bioinformatics Intervention in Elucidating Structural and Functional Attributes of Plant Specific Transcription Factors: A Review

Yadav Dinesh, Malviya Neha, Nasim Jeya and Kumar Rajendra

Innovations in sequencing technologies have led to the deciphering of several plant genome sequences in the recent years, resulting in a substantial expansion of sequence databases. The developments of web based bioinformatics tools are playing a significant role in the analysis of whole genome sequences to get an insight into the complexity of gene expression and regulation in plants. Transcription factors (TF) are known to be important element associated with gene regulation by interacting with specific sequences of promoters of the concerned genes. The importance of TF in gene expression and regulation was realized based on the fact that 5-10% of whole genome sequence represents genes coding for TFs. There are several types of TFs, some are common in plants and animals while there exist plant specific TFs known to be associated with functions influencing growth and development of plants. The presence of specific type of DNA binding domain in the TFs is considered to be one important criterion for classification of TFs. Bioinformatics based assessment of these plants specific TFs gained momentum with the development of crop specific TF databases, freely available to the researchers. The reports of genome-wide in-silico prediction, bioinformatics based sequence characterization, wet-lab based cloning and expression profiling of several plant specific TFs, representing crops whose genome sequences have been deciphered, is substantially increasing. The potential of stress responsive TFs in developing biotic or abiotic tolerant crops by transgenic approach have been realized by the plant biotechnologists and are being investigated extensively.

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Bruchid resistance in food legumes-an overview

Tripathy Swapan K.

Bruchids are the most notorious pest that can devastate the entire seed lots of food legumes in storage. The extent of damage varied with legume crops and bruchid spp. Several options including insecticidal pesticide application are currently available to check bruchid infestation. However, development of cultivar with adequate level of host plant resistance can be cost effective, durable and eco-friendly approach. In this pursuit, the author presented a detailed review of the insect life style, screening technique, source and mechanisms of resistance, mode of inheritance and novel breeding strategies including genetic transformation and use of molecular and seed protein markers in marker aided selection.

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