Research Journal of Biotechnology

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Research Journal of Biotechnology

Seed storage protein profiling and phylogenetic relationships of Capsicum annum L. cultivars using SDS-PAGE

Peddakasim D., Suneetha P., Lakshmi Sahithya U., Srideepthi R. and Krishna M. S. R.

Capsicum annuum L. is a commercial crop which belongs to the Solanaceae family and it is widely grown in tropical and sub tropical regions of the world. Capsicum seeds consist of different types of soluble proteins. In the present study we analyzed the biochemical relationship among the sixteen genotypes of Capsicum annum L. Seed protein profiling was generated in sixteen genotypes of chilli seeds through SDS- PAGE. The analysis showed 92 bands and considerable variation in banding pattern. Dendrogram was constructed based on protein profiling score using NTSYS software. Total sixteen genotypes were grouped into four clusters. Based on this diversity study, CA 1, 2 and 5 are highly diversified with CA 12, 13, 15 and 16 genotypes. Based on this study, highly diversified genotypes could be utilised for chilli breeding programme.

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Study on the usage of aquatic vegetation and combined feed in certain Cyprinids species feeding

Gîlcă Valerica, Gîlcă Ioan and Radu Rusu Răzvan

The trial aimed to assess the weight gain of four cyprinids species: Cyprinus carpio (common carp), Ctenopharyngodon idella (grass carp), Hypophthalmichthys molitrix (silver carp) and Aristichtys nobilis (bighead carp) when natural feed resources and combined feed were used comparatively. The experiment was developed during the 2013 vegetative period on the four species hosted within the Aquaculture and Aquatic Ecology Scientific Station throughout 45 days (20 July – 03 September). During the research, main physical and chemical water traits (temperature, transparency, color, pH, oxygen, calcium, magnesium, chlorides) from two fishery pools were analyzed. Literature standardized methods were applied for each parameter assessment9,10,14,16,17. Variation limits for the physical and chemical water traits did not negatively affect production in the studied pools. At 45 days after experiment onset, the best results related to the average body weight were achieved by the LE group (supplementary fed with combined feed). Thus, for the specimens fed with concentrated feed, compared with those fed on natural basis only, the body weights were higher: Aristichtys nobilis (bighead carp) was 21.5% heavier; Hypophthalmichthys molitrix (silver carp) and Ctenopharyngodon idella (grass carp) were 27.5 % heavier and Cyprinus carpio (common carp) was 29.0% better developed.

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Isolation and screening of Aspergillus species for protease production

Tiwari K. L. and Rizvi A. F.

Proteases are group of enzymes which hydrolyze protein. Proteases have long been used in processes such as brewing, baking, meat tenderization and detergent additives. Present research work was carried out with an aim to isolate Aspergillus species from the soil of Chhattisgarh region and to evaluate their potential for alkaline protease and neutral protease production. Total 15 Aspergillus species were isolated from soil and only five Aspergillus species showed positive results for proteases. Extracellular and intracellular neutral protease activity was high in A. luchensis. Alkaline protease activity was also estimated from all five species. Extracellular and intracellular alkaline protease activities were tested separately. Maximum extracellular and intracellular alkaline protease activity was observed by A. nidulans.

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Encryption for Sharing of Secret Medical Images

Sangeetha M., Arumugam C. and Senthilkumar K. M.

Visual Cryptography Scheme (VCS) is a type of secret sharing scheme which allows the encoding of a secret image into n shares distributed to n participants. Each share constitutes some information and when k shares out of n stack together the secret will reveal. However; less than k shares will not work. The advantage of the visual secret sharing scheme is its decryption process i.e. to decrypt the secret using Human Visual System without any computation. Traditional Visual Cryptography suffers from share identification problem. This problem can be solved by Multiple-Secret threshold visual cryptography (MVCS) which adds a meaningful cover image in each share. The proposed work presents threshold Visual cryptographic schemes in Color Images. This method uses half toning method to provide color image as a secret image. Then the secret image can be embedded in the original image by generating shares using Zigzags scanning method. Experimental result of proposed system provides robust security than conventional visual cryptographic schemes.

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Biological Synthesis of Silver Nanoparticles using Colocasia Extract and their antimicrobial activity

Boruah Himangshu, Talukdar Binita, Parveen Assma, Goswami Gunajit, Barooah Madhumita and Boro Robin Chandra

Silver nanoparticles were synthesized by a simple, non-toxic, eco-friendly biological reduction method using colocasia extract as reducing agent. A variety of nanoparticles were formed when the concentration of the reducing agent was increased with respect to the silver nitrate solution. The reaction mixtures displayed variation in colours and characteristic UV-VIS spectra of silver nanoparticles with the increasing concentration of colocasia extract. A single SPR band in between 419 to 438 nm in UV-VIS spectroscopy revealed the formation of silver nanoparticles. TEM analysis of the silver nanoparticles confirmed the spherical shape and size distribution of the nanoparticles found in the range of 10-30nm. Antibacterial activity of the Ag-NPs was confirmed by disk diffusion method against Bacillus flexus, Escherichia coli, Serratia marcescens and Staphylococcus aureus.

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Characterization and expression of PGK2 and PHKG2 in testis tissues from fertile and sterile Banna Mini-pig Inbred Line (BMI)

Wang Pei, Wang Shuyan, Qin Caiyan and Huo Jinlong

The objective of this study was to clone BMI sterile and fertile PGK2 and PHKG2 genes and analyze the expression patterns in the different reproduction’s testis of Banna mini-pig inbred line. Using RT-PCR, PGK2 and PHKG2 cDNAs were cloned and bioinformatics analysis was performed. Using GAPDH gene as a control, the expressions of PGK2 and PHKG2 mRNA in the testis of sterile and fertile BMI boars were detected by semi-quantitative methods. The full-length coding sequence (CDS) of the BMI PGK2 consists of 1254 bp which encodes a 417 amino acid protein with molecular mass of 44.84 kD and a pI of 8.31. And the PHKG2 CDS is 1221 bp in length which encodes a 406 amino acid protein with molecular mass of 46.56 kD and a pI of 5.86. Four and eight nucleotide differences were found in the cDNAs of PGK2 and PHKG2 between BMI but only one missing nucleotide mutation at c.970 (T>A) changes aspartic acid (Asn) at p.324 in BMI fertile boars to tyrosine (Tyr) in the BMI sterile boars protein. The putative proteins of BMI PGK2 and PHKG2 both located in the cytoplasm with high reliability contained the conserved domain and active sites of protein kinase superfamily and no signal peptide. Moreover, there is a coiled-coil motif in the N terminus of BMI PHKG2. Phylogenetic analyses based on the PGK2 and PHKG2 amino acid sequences showed that BMI was grouped with cattle, sheep and horse. The RT-PCR analysis showed that the expression of PGK2 and PHKG2 was similar in the testes of fertile and sterile BMI boar. These data provide the primary foundation for further insights into the BMI PGK2 and PHKG2 genes.

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Isolation of a unique Phenol degrading bacterial strain Escherichia coli moh 1 from effluent of an edible oil industry in Chennai, India

Fayidh M. A., Kalleary Sabina, Kasirajan Sudharsan and Muthusamy Sukumar

A novel environmental bacterial strain, Escherichia coli moh1 was isolated and characterized from oil industry effluent in Chennai, Tamil Nadu, India. Primary growth experiments showed that this strain has the ability to grow in presence of up to 800 mg/l of phenol in NB medium at 37°C with the initial inoculum of ~104 CFU. The degradation rate of phenol (200 mg/l) was checked through monitoring growth at 24h using the HPLC system. The results showed considerable decrease in phenol concentration which was obvious from the decrease in peak intensity, a distinct change in the retention time and peak profile indicating complete degradation of phenol. 16SrRNA sequence of the isolate was deposited to GenBank at NCBI and registered as a unique phenol degrading strain.

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Production and purification of peroxidase from horseradish (Armoracia rusticana) roots using ultrafiltration

Mei Qin

A simple and efficient procedure for the large-scale production of peroxidase from horseradish (Armoracia rusticana) roots using only ultrafiltration has been developed. After two-stage ultrafiltration process, the enzyme was purified 238-fold with a specific activity of 642.6 U/mg protein and a yield of 86 %. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed a main band near 41 kDa. The purified peroxidase had an alpha-helix content of 54.7% consistent with the theoretical value. This shows that the purified peroxidase folded with a reasonable secondary structure.

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Relationships between endogenous hormonal content and direct somatic embryogenesis in Watermelon (Citrullus lanatus) cotyledons

Hui Jun Zhang and Qing Wang

Cotyledons of Watermelon (Citrullus lanatus) were cultured in vitro on medium deprived of plant growth regulators. Inbred lines varying in their embryogenic capacity were studied after 90 days in culture media. Endogenous levels of indole-3-acetic acid (IAA), abscisic acid (ABA) and trans-zeatin riboside (ZR) were analyzed in embryogenic cotyledons. Significant differences were observed in total ABA, IAA, ZR, concentrations of the embryogenic cotyledons. On the contrary, lower Z and ACC contents and also a reduced balance between Z and IAA levels were related with the embryogenic capacity of the cotyledons. These results suggest that the difference in somatic embryo formation capacity observed between embryogenic cotyledons is related to their endogenous Z contents and that the endogenous hormonal balance between Z and IAA is an important index defining the embryogenic potential in peach cotyledons.

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Purification and further characterizations of the cellulase from the spent mushroom compost of Hypsizygus marmoreus

Yunlong Yang, Caimei Huang and Kaihui Hu

Using the orthogonal experimental design method involving three factors and four levels, the extraction conditions of cellulase from the spent mushroom compost of Hypsizygus marmoreus were studied and the optimal extraction conditons were determined through orthogonal range and variance analysis. Then, through the combination of sulfate ammonium precipation, sephadex G100 gel filtration and DEAE C-52 ion exchange chromatography, the cellulase was puritfied to homogeneitiy. Results showed that this cellulase was purified up to 12.13 fold with a specific activity of 1121.82 U/mg and proved to be a monomeric enzyme with a molecular mass of 27.8 kDa. The enzyme displayed its optimal activiy at 500C and pH 5.0 and possessed “wide-stable” nature due to its stability in the pH range of 3.0 to 10.0. The enzyme got inhibited by Cu2+, Zn2+ and Fe3+ but activated by Mg2+ and Mn2+.

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Molecular characterization and expression pattern of tobacco (Nicotiana tabacum) TPX2 peroxidase gene

Xu Shengguang, Su Yuan and Liu Yonggang

The complete mRNA sequence of one tobacco (nicotiana tabacum) gene-thioredoxin-dependent peroxidase 2 (TPX2 peroxidase) was amplified using the rapid amplification of cDNA ends methods. The full-length tobacco TPX2 peroxidase gene mRNA was 1,269bp containing an 987 bp open reading frame which encodes a protein of 328 amino acids. Sequence analysis revealed that the TPX2 peroxidase of tobacco shares high homology with the TPX2 peroxidase of lycopersicon esculentum (81%), sweet potato (75%), wine grape (73%), eucommia ulmoides (73%) and cacao (70%). Results also showed that tobacco TPX2 peroxidase gene has a closer genetic relationship with the TPX2 peroxidase gene of lycopersicon esculentum. The expression profile was studied and the results indicated that tobacco TPX2 peroxidase gene was highly expressed in root but hardly expressed in leaf, stem and flower.

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Survey of Radiology Clinical Methodologies for Breast Cancer’s Symptoms Detection

Sakthisudhan K., Saravana Kumar N. and Thangaraj P.

Breast cancer is a major challenge and health issue of the present society. The death rate has doubly risen in the past 30 years. The various diagnosis systems are authorized and followed to reduce the death rate. The Magnetic Resonance Imaging (MRI) and Positron Emission Tomography (PET) are existing diagnosis trials. The X-ray mammography (XRM) is an efficient diagnostic tool but it has some limitations in the screening detection methods. They are, the heavy chemical dose applied on the human breast of body to affect ionization effects within the body and harmful radiation effects during the clinical tests. Both of the above results provide the injurious health issues. Normally, these radiology diagnostic trials do not offer the tissue’s sample based analyzed report. These drawbacks of MRI, XRM and other existing techniques are recovered using the Microwave Imaging (MI) methods. The available diagnostic methods (Magnetic resonance imaging, Microwave Radiometry) are studied and compared with microwave imaging method is presented in this survey.

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