Bulani, SMoleleki, LAlbertyn, JMoleleki, J2013-04-222013-04-222012-05Bulani, S.I, Moleleki, L, Albertyn, J and Moleleki, N. 2012. Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica. AMB Express, vol. 2(27), pp 1-82191-0855http://www.amb-express.com/content/2/1/27http://hdl.handle.net/10204/6709Copyright: 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 1-8In 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.enmCherryrDNA vectorYlCWP1Yarrowia lipolyticaCell surface displayArticleBulani, S., Moleleki, L., Albertyn, J., & Moleleki, J. (2012). Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica. http://hdl.handle.net/10204/6709Bulani, S, L Moleleki, J Albertyn, and J Moleleki "Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica." (2012) http://hdl.handle.net/10204/6709Bulani S, Moleleki L, Albertyn J, Moleleki J. Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica. 2012; http://hdl.handle.net/10204/6709.TY - Article AU - Bulani, S AU - Moleleki, L AU - Albertyn, J AU - Moleleki, J AB - 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. DA - 2012-05 DB - ResearchSpace DP - CSIR KW - mCherry KW - rDNA vector KW - YlCWP1 KW - Yarrowia lipolytica KW - Cell surface display LK - https://researchspace.csir.co.za PY - 2012 SM - 2191-0855 T1 - Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica TI - Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica UR - http://hdl.handle.net/10204/6709 ER -