| Title |
Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica
|
|---|---|
| Published in |
AMB Express, May 2012
|
| DOI | 10.1186/2191-0855-2-27 |
| Pubmed ID | |
| Authors |
Siyavuya Ishmael Bulani, Siyavuya Ishmael Bulani, Lucy Moleleki, Jacobus Albertyn, Ntsane Moleleki |
| 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. |
Mendeley readers
The data shown below were compiled from readership statistics for 32 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| France | 1 | 3% |
| South Africa | 1 | 3% |
| Unknown | 30 | 94% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 7 | 22% |
| Researcher | 7 | 22% |
| Student > Master | 3 | 9% |
| Student > Doctoral Student | 2 | 6% |
| Professor | 2 | 6% |
| Other | 3 | 9% |
| Unknown | 8 | 25% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 12 | 38% |
| Biochemistry, Genetics and Molecular Biology | 8 | 25% |
| Immunology and Microbiology | 1 | 3% |
| Chemistry | 1 | 3% |
| Engineering | 1 | 3% |
| Other | 0 | 0% |
| Unknown | 9 | 28% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 20 May 2012.
All research outputs
#20,157,329
of 22,665,794 outputs
Outputs from AMB Express
#963
of 1,227 outputs
Outputs of similar age
#147,861
of 163,617 outputs
Outputs of similar age from AMB Express
#3
of 5 outputs
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So far Altmetric has tracked 1,227 research outputs from this source. They receive a mean Attention Score of 2.8. This one is in the 1st percentile – i.e., 1% of its peers scored the same or lower than it.
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