Open Access Original Research Article

Insilico Prediction of T-cell Epitopes to Therapeutic Interferon -Beta (IFN-β) Protein

Swathi Krishna Reddy, Venkata Bharat Kumar Pinnelli

Biotechnology Journal International, Page 1-23
DOI: 10.9734/BJI/2016/29184

Aims: Several studies have reported the existence for T helper cell epitopes with the persistence of unwanted immune reactions for several protein drugs. T-cell epitope is an amino acid or set of amino acids that are capable of being recognized form one or more T-cell receptors. There is also an indication that T helper cells are involved in the anti-drug antibodies development to therapeutic interferon beta-1a. Protein drugs containing Major histocompatibility complex class II T cell epitopes are likely to elicit anti-drug antibodies. Binding specificity between T-cell epitopes and major histocompatibility molecules are the most important determinant step in finding the T-cellular immune responses. The data obtained from the present study provides new insights into prediction of therapeutic Interferon beta T helper cells epitopes using T cell epitope prediction tools, mapping of clusters of predicted epitopes.

Study Design: Insilico analysis by bioinformatics tools was to predict T-cell epitopes of Interferon beta-1a.

Methodology: Several Insilico prediction tools (immunoinformatics tools) including Proped, NetMHCIIpan3.0 and Immune Epitope Database Analysis Resource (IEDB-AR) are available to map the potential major histocompatibility class II T cell epitopes. After predicting potential T-cell epitopes, epitopes were mapped on interferon beta-1a using MIMOX2 server.

Results: The potential MHC class II immunogenic sequence of 50 amino acids “TRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTIVRVEILRNFYFINRLTG” With IFN-β-1a (position 111-161) were identified. This study can provide the understanding the relevance to T-cell activation for prediction and assessment of unwanted immune responses.

Conclusions: Insilico prediction by using the available tools helps in reducing the time and cost for the immunologists during the vaccine design. By predicting them we will come to know, which peptides play major role and synthesize them using invitro technologies.

Open Access Original Research Article

Plant Regeneration and Its Functional Analysis within Transgenic Rice of ASG-1, an Apomixis-specific Gene Isolated from Apomictic Guinea Grass

Lanzhuang Chen, Liming Guan, Daiki Toyomoto, Toru Sugita, Takuro Hamaguchi, Reiji Okabe

Biotechnology Journal International, Page 1-13
DOI: 10.9734/BJI/2016/29146

Aims: To analyze the function of the ASG-1, an apomixis-specific gene has been isolated from apomictic guinea grass, the combination of Agrobacterium-mediated transformation system and pSMA35H2-NG binary vector were used for transformation of ASG-1 to ‘Nipponbare’ of rice, and its functional analysis of transgenic rice was carried out.

Study Design: 1) plant regeneration of transgenic rice of ASG-1 was achieved, and 2) it was followed by experiments with the obtained non-graining ear in the whole plants (NGEP) using comparative observation in both pollens of guinea grass and the T0 NGEP with Nomarski differential interference-contrast microscopy (DIC).

Place and Duration of Study: Faculty of Environmental and Horticultural Science, Minami Kyushu University, between September 2012 and October 2015.

Methodology: As a preliminary transformation, a β-glucuronidase (GUS) was used to infect the callus. Plant regeneration of the transgenic rice of ASG-1 were achieved. DNAs of the transgenic rice were used for detection of ASG-1 by PCR. For analysis of NGEP, the anthers and ovaries of the transgenic rice and guinea grass were used for comparative observation with DIC.

Results: 1) GUS was successfully expressed in calli of both rice and guinea grass. 2) The transgenic rice plants of ASG-1 were then obtained from the culture of seed-derived calli. 3) The natures of T0 callus and T1 plants were confirmed. 4) A subset (~15%) of T0 plants showed morphological sterility with NGEP. 5) The ovaries of the NGEP showed identical characteristics to “Nipponbare” and other normal T1 plants; however, the pollen showed normal rates of <38%. 6) The pollen of guinea grass at anthesis showed rates of normal pollen as low as that of the NGEP rice.

Conclusion: ASG-1 may play a role in regulating pollen sterility. Comparing these findings with our previous reports, we identified co-localization for ASG-1 in pollen of both rice and guinea grass, suggesting that the trait of ASG-1 influencing pollen sterility might be inherited from guinea grass to rice through the transformation process.

Open Access Original Research Article

Assessing the Genotoxic Effects of Aqueous Leaf Extract of Neem (Azadirachta indica A. Juss), Cosmetics and Alomo Bitters (Alcoholic Drink) Using Allium Test

G. C. Obute, C. Ekeke, U. A. Nwagwu

Biotechnology Journal International, Page 1-13
DOI: 10.9734/BJI/2016/29042

Genotoxic effect of aqueous leaf extract of Neem (Azadirachta indica) NZI, Alomo bitters alcoholic drink (ABAD) and cosmetics (Baby-face facial cleanser, BFFC; Peau clair body lotion, PCBL and Vanilla essence roll-on, VERO) were assessed using Allium test. The root tips were treated with serial dilutions of the test chemicals at 20%, 40%, 60% and 100% for 3 hours along with the control, hydrolyzed in 18% HCl, squashed and stained with FLP orcein. Aberrations such as C-metaphase, precocious, stickiness (sticky metaphase), ghost cells, nuclear lesions, chromosome erosion, laggards, binucleated cells, anaphase bridges and vagrant chromosomes were observed in the root tips studied. These products are used widely by humans around the world and Nigeria in particular. Although, the products elicited varying chromosomal abnormalities, they also have beneficial effects. Also, the products showed genotoxic effects even at low concentrations on Allium cepa root cells. In a long run, the use of these products may cause serious damages on the chromosome of plants, animals and humans. More effort is required to actually ascertain the active constituents of these products that elicited the genotoxic effects. Constant use and consumption of these products is therefore discouraged.

Open Access Original Research Article

Molecular Characterization of Orobanche crenata in Egypt Using ISSR Markers and Its Relation to Faba Bean Breeding

M. M. F. Abdalla, M. M. Shafik, Sabah M. Attia, Hend A. Ghannam

Biotechnology Journal International, Page 1-11
DOI: 10.9734/BJI/2016/27216

Orobanche crenata is an annual parasitic weed that causes heavy losses to its host crop faba bean (Vicia faba). Determining the genetic diversity in Orobanche germplasm is a preliminary crucial step in faba bean breeding.It helps in identifying liable criteria of host tolerance. The current study aimed to determine the genetic diversity for Orobanche collected from two divergent locations in Egypt (Giza and Sids). The inter simple sequence repeats (ISSR) markers were used to determine genetic relationships among and within the collected groups. The number of produced fragments covered 218 to 980 bp of the total of Orobanche crenata genome. The five used primers generated a total of 73 ISSR products with an average of 14.6 products / primer. The polymorphic information content (PIC) ranged from 0.86- 0.94.

Analyses of molecular variance indicated significant difference within each region. The difference between the two locations was not significant (3%) due to great variation within locations (97%). The cluster analysis devided the 96 Orobanche samples into five sub-clusters. The study signified the efficiency of ISSR molecular technique in estimating genetic relationships across the two divergent locations at the genomic level. It also was consistence with the predominantly allogamous behaviour of O. crenata. Because of Orobanche diversity, faba bean breeders should develop host varieties with heterogeneous background and don‟t release any pure line host in infested soils.

Open Access Review Article

Transgenic Cotton Development: Agro-economic Analysis and Comparative Study

Julian Witjaksono, Asmin .

Biotechnology Journal International, Page 1-15
DOI: 10.9734/BJI/2016/29603

The development of transgenic crop since its first launched to the public in 1995 results in high expectation in order to boost the agricultural productivity, particularly in cotton. Higher yield and higher return are the expectation of cotton growers especially for poor-resource farmers due to low income household. This study provides the evidence of implementing Genetically Modified (GM) cotton based on the meta-data which derived from indvidual studies more than one decade in China, India, USA and Australia as the comparative study. Economic performance is the analysis of economic indicators such as yield gain, seed cost, pesticide cost, management and labor cost, and net return in which the comparison between GM cotton and its counterpart worldwide overtime. Study findings that it is clear that this technology is not superior and still need to be suitable for the given production situation, and also depending on the specific pest pressure and other relevant local condition to optimize per hectare returns. This study results that this merit technology can vary in different ecological environments.