Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3

Genetic Engineering Related Abstracts

3 Constitutive Flo1p Expression on Strains Bearing Deletions in Genes Involved in Cell Wall Biogenesis

Authors: Lethukuthula Ngobese, Abin Gupthar, Patrick Govender

Abstract:

The ability of yeast cell wall-derived mannoproteins (glycoproteins) to positively contribute to oenological properties has been a key factor that stimulates research initiatives into these industrially important glycoproteins. In addition, and from a fundamental research perspective, yeast cell wall glycoproteins are involved in a wide range of biological interactions. To date, and to the best of our knowledge, our understanding of the fine molecular structure of these mannoproteins is fairly limited. Generally, the amino acid sequences of their protein moieties have been established from structural and functional analysis of the genomic sequence of these yeasts whilst far less information is available on the glycosyl moieties of these mannoproteins. A novel strategy was devised in this study that entails the genetic engineering of yeast strains that over-express and release cell wall-associated glycoproteins into the liquid growth medium. To this end, the Flo1p mannoprotein was overexpressed in Saccharomyces cerevisiae laboratory strains bearing a specific deletion in KNR4 and GPI7 genes involved in cell wall biosynthesis that have been previously shown to extracellularly hyper-secrete cell wall-associated glycoproteins. A polymerase chain reaction (PCR) -based cloning strategy was employed to generate transgenic yeast strains in which the native cell wall FLO1 glycoprotein-encoding gene is brought under transcriptional control of the constitutive PGK1 promoter. The modified Helm’s flocculation assay was employed to assess flocculation intensities of a Flo1p over-expressing wild type and deletion mutant as an indirect measure of their abilities to release the desired mannoprotein. The flocculation intensities of the transformed strains were assessed and all the strains showed similar intensities (>98% flocculation). To assess if mannoproteins were released into the growth medium, the supernatant of each strain was subjected to the BCA protein assay and the transformed Δknr4 strain showed a considerable increase in protein levels. This study has the potential to produce mannoproteins in sufficient quantities that may be employed in future investigations to understand their molecular structures and mechanisms of interaction to the benefit of both fundamental and industrial applications.

Keywords: Genetic Engineering, Flocculation, glycoproteins, over-expression

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2 Sensing Endocrine Disrupting Chemicals by Virus-Based Structural Colour Nanostructure

Authors: Lee Yujin, Han Jiye, Oh Jin-Woo

Abstract:

The adverse effects of endocrine disrupting chemicals (EDCs) has attracted considerable public interests. The benzene-like EDCs structure mimics the mechanisms of hormones naturally occurring in vivo, and alters physiological function of the endocrine system. Although, some of the most representative EDCs such as polychlorinated biphenyls (PCBs) and phthalates compounds already have been prohibited to produce and use in many countries, however, PCBs and phthalates in plastic products as flame retardant and plasticizer are still circulated nowadays. EDCs can be released from products while using and discarding, and it causes serious environmental and health issues. Here, we developed virus-based structurally coloured nanostructure that can detect minute EDCs concentration sensitively and selectively. These structurally coloured nanostructure exhibits characteristic angel-independent colors due to the regular virus bundle structure formation through simple pulling technique. The designed number of different colour bands can be formed through controlling concentration of virus solution and pulling speed. The virus, M-13 bacteriophage, was genetically engineered to react with specific ECDs, typically PCBs and phthalates. M-13 bacteriophage surface (pVIII major coat protein) was decorated with benzene derivative binding peptides (WHW) through phage library method. In the initial assessment, virus-based color sensor was exposed to several organic chemicals including benzene, toluene, phenol, chlorobenzene, and phthalic anhydride. Along with the selectivity evaluation of virus-based colour sensor, it also been tested for sensitivity. 10 to 300 ppm of phthalic anhydride and chlorobenzene were detected by colour sensor, and showed the significant sensitivity with about 90 of dissociation constant. Noteworthy, all measurements were analyzed through principal component analysis (PCA) and linear discrimination analysis (LDA), and exhibited clear discrimination ability upon exposure to 2 categories of EDCs (PCBs and phthalates). Because of its easy fabrication, high sensitivity, and the superior selectivity, M-13 bacteriophage-based color sensor could be a simple and reliable portable sensing system for environmental monitoring, healthcare, social security, and so on.

Keywords: Genetic Engineering, M-13 bacteriophage, colour sensor, EDCs

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1 Problems and Prospects of Agricultural Biotechnology in Nigeria’s Developing Economy

Authors: Samson Abayomi Olasoju, Olufemi Adekunle, Titilope Edun, Johnson Owoseni

Abstract:

Science offers opportunities for revolutionizing human activities, enriched by input from scientific research and technology. Biotechnology is a major force for development in developing countries such as Nigeria. It is found to contribute to solving human problems like water and food insecurity that impede national development and threaten peace wherever it is applied. This review identified the problems of agricultural biotechnology in Nigeria. On the part of rural farmers, there is a lack of adequate knowledge or awareness of biotechnology despite the fact that they constitute the bulk of Nigerian farmers. On part of the government, the problems include: lack of adequate implementation of government policy on bio-safety and genetically modified products, inadequate funding of education as well as research and development of products related to biotechnology. Other problems include: inadequate infrastructures (including laboratory), poor funding and lack of national strategies needed for development and running of agricultural biotechnology. In spite of all the challenges associated with agricultural biotechnology, its prospects still remain great if Nigeria is to meet with the food needs of the country’s ever increasing population. The introduction of genetically engineered products will lead to the high productivity needed for commercialization and food security. Insect, virus and other related diseases resistant crops and livestock are another viable area of contribution of biotechnology to agricultural production. In conclusion, agricultural biotechnology will not only ensure food security, but, in addition, will ensure that the local farmers utilize appropriate technology needed for large production, leading to the prosperity of the farmers and national economic growth, provided government plays its role of adequate funding and good policy implementation.

Keywords: Biotechnology, Food Security, Biosafety, Genetic Engineering, Genetic Modification

Procedia PDF Downloads 24