http://hdl.handle.net/10204/893 2013-06-19T23:40:35Z http://hdl.handle.net/10204/6709 Title: Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica Authors: Bulani, S; Moleleki, L; Albertyn, J; Moleleki, J Abstract: In this study, a novel rDNA based plasmid was developed for display of heterologous proteins on the cell surface of Yarrowia lipolytica using the C- terminal end of the glycosylphosphatidylinositol (GPI) anchored Y. lipolytica cell wall protein 1 (YlCWP1). mCherry was used as a model protein to assess the efficiency of the constructed plasmid. Y. lipolytica transformants harbouring the expression cassettes showed a purple colour phenotype on selective YNB-casamino plates as compared to control cells indicating that mCherry was displayed on the cells. Expression of mCherry on cells of Y. lipolytica was confirmed by both fluorescent microscopy and flow cytometry. Furthermore, SDS-PAGE analysis and matrix-assisted laser desorption/ionization (MALDI)-time-of (TOF)-mass spectrometry (MS) peptide mass fingerprinting (PMF) confirmed that the protein cleaved from the yeast cells using enterokinase was mCherry. Efficient cleavage of mCherry reported in this work offers an alternative purification method for displayed heterologous proteins on Y. lipolytica cells using the plasmid constructed in this study. The developed displaying system offers great potential for industrial production and purification of heterologous proteins at low cost. Description: Copyright: 2012 BioMed Central. This is an Open Access journal. This journal authorizes the publication of the information herewith contained. Published in AMB Express, vol. 2(27), pp 1-8 2012-05-01T00:00:00Z http://hdl.handle.net/10204/5414 Title: PALM and STORM: unlocking live-cell super-resolution Authors: Henriques, R; Griffiths, C; Rego, EH; Mhlanga, MM Abstract: Live-cell fluorescence light microscopy has emerged as an important tool in the study of cellular biology. The development of fluorescent markers in parallel with super-resolution imaging systems has pushed light microscopy into the realm of molecular visualization at the nanometer scale. Resolutions previously only attained with electron microscopes are now within the grasp of light microscopes. However, until recently, live-cell imaging approaches have eluded super-resolution microscopy, hampering it from reaching its full potential for revealing the dynamic interactions in biology occurring at the single molecule level. Here we examine recent advances in the super-resolution imaging of living cells by reviewing recent breakthroughs in single molecule localization microscopy methods such as PALM and STORM to achieve this important goal. Description: Copyright: 2011 Wiley Blackwell. This is an ABSTRACT ONLY 2011-05-01T00:00:00Z http://hdl.handle.net/10204/4457 Title: Effect of expressing an anti-HIV lectin, Griffithsin, in different plant cellular compartments Authors: Stark, T; Chakauya, E; Kabamba, A; Morris, L; Rybicki, E; Chikwamba, R Abstract: Sub-Saharan Africa remains the region most heavily affected by HIV, and the majority of new infections from heterosexual contact are in women. Griffithsin (GRFT), a lectin isolated from the red algae Griffithsia, has shown extreme potency against the Human Immunodeficiency Virus (HIV) in vitro in very low quantities. The molecular target of GRFT is the high mannose residues on the viral gp120 coat protein. Low cost production of GRFT and other anti-HIV drugs at minimal costs is crucial seeing that the high demand for anti-HIV drugs reside in poor developing countries. Description: CSIR 3rd Biennial Conference. Science Real and Relevant, CSIR International Convention Centre, Pretoria, 30 August – 1 September 2010 2010-08-01T00:00:00Z http://hdl.handle.net/10204/4169 Title: Morphological variation of sorghum landraces from semi-arid areas of Zimbabwe Authors: Mujaju, C; Chakauya, E Abstract: Sorghum (Sorghum bicolor) is one of the most important cereal crops cultivated in, the semi-arid areas of Zimbabwe where persistent drought, high temperatures and poor edaphic conditions are endemic. Because of their adaptation, landraces are the mainstay of smallholder agriculture in such areas. Unfortunately, limited studies have been done to quantify the genetic diversity of this material in order to formulate the necessary conservation strategies. This study analysed the genetic diversity of 47 sorghum landraces from two districts of Zimbabwe, Nyanga North and Tsholotsho using 24 Sorghum agromorphological descriptors. Genetic similarities (Manhattan coefficient) were calculated and genetic relationships between accessions were analysed by principal component analysis and cluster analysis. The landraces grouped into six clusters according to the geographical location where they were collected from, suggesting environmental adaptation. Accessions with the same name had a tendency to group together, although some material was found scattered throughout the dendrogram. Agromorphological traits were highly variable even for landraces with the same farmer-given name and source. The results also alluded to the heterogeneity of farmer varieties, whose naming appears to be a function of a few traits. The study suggests that optimisation of on-farm conservation strategy for this germplasm should primarily focus on high diversity areas and perhaps recognise those traits envisaged to be of importance by farmers for varietal identification Description: Copyright: 2008 Asian Network for Scientific Information 2008-01-01T00:00:00Z