Genome Evolution in Yeasts

Dujon, Bernard

doi:10.1002/9780470015902.a0023986

Cited by 175 publications

(262 citation statements)

References 98 publications

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“…Yet, the genetic tractability of Y. lipolytica [18][19][20][21][22][23][24] coupled with its modest, innate de novo lipogenesis (B10-15% lipid content in wild type 13,14,[25][26][27] ) make it a potential candidate as a platform organism for superior lipid production.…”

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confidence: 99%

Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production

et al. 2014

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Economic feasibility of biosynthetic fuel and chemical production hinges upon harnessing metabolism to achieve high titre and yield. Here we report a thorough genotypic and phenotypic optimization of an oleaginous organism to create a strain with significant lipogenesis capability. Specifically, we rewire Yarrowia lipolytica's native metabolism for superior de novo lipogenesis by coupling combinatorial multiplexing of lipogenesis targets with phenotypic induction. We further complete direct conversion of lipid content into biodiesel. Tri-level metabolic control results in saturated cells containing upwards of 90% lipid content and titres exceeding 25 g l À 1 lipids, which represents a 60-fold improvement over parental strain and conditions. Through this rewiring effort, we advance fundamental understanding of lipogenesis, demonstrate non-canonical environmental and intracellular stimuli and uncouple lipogenesis from nitrogen starvation. The high titres and carbon-source independent nature of this lipogenesis in Y. lipolytica highlight the potential of this organism as a platform for efficient oleochemical production.

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confidence: 99%

Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production

et al. 2014

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“…According to Dujonet al, (2004), the big compensation gap can reduce the agancy cost efficiently, which is important to motivate executives and staff to work hard in order to get a better promotion [8]. Based on the perspective I mentioned above, the first hypothesis is that:…”

Section: Hypothesis Developmentmentioning

confidence: 99%

Research on the Relationship between Compensation Gaps of Executive-staff and Corporate Performance

Han¹,

Huangfu²

2017

dtssehs

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Abstract. This paper aims at exploring the relationship between the corporate performance and compensation gaps of executive-staff, by using the samples of Chinese firms from Shanghai Stock Exchange over the period of 2005-2015. We measured the corporate performance by computing the return on equity. And we also used the average compensation of the top three executives divided by the average of the ordinary employees to quantify the compensation gap. Ultimately, we conclude that there is a significant "inverse U" relationship between the compensation gap and the corporate performance.

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“…The C. glabrata SIR3 destination vector was constructed by PCR-amplifying the regions flanking the SIR3 ORF from strain CBS138 (Dujon et al 2004). Primers 3220 and 3221 were used to amplify the 59 flanking region; primers 3222 and 3223 were used to amplify the 39 flanking region.…”

Section: Construction Of C Glabrata Sir3 Gateway Vectorsmentioning

confidence: 99%

Avoiding the Ends: Internal Epitope Tagging of Proteins Using Transposon Tn7

et al. 2015

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Peptide tags fused to proteins are used in a variety of applications, including as affinity tags for purification, epitope tags for immunodetection, or fluorescent protein tags for visualization. However, the peptide tags can disrupt the target protein function. When function is disrupted by fusing a peptide to either the N or C terminus of the protein of interest, identifying alternative ways to create functional tagged fusion proteins can be difficult. Here, we describe a method to introduce protein tags internal to the coding sequence of a target protein. The method employs in vitro Tn7-transposon mutagenesis of plasmids for random introduction of the tag, followed by subsequent Gateway cloning steps to isolate alleles with mutations in the coding sequence of the target gene. The Tn7-epitope cassette is designed such that essentially all of the transposon is removed through restriction enzyme digestion, leaving only the protein tag at diverse sites internal to the ORF. We describe the use of this system to generate a panel of internally epitope-tagged versions of the Saccharomyces cerevisiae GPI-linked membrane protein Dcw1 and the Candida glabrata transcriptional regulator Sir3. This internal protein tagging system is, in principle, adaptable to tag proteins in any organism for which Gateway-adapted expression vectors exist. KEYWORDS Saccharomyces; epitope tag; protein tagging; transposition R ECOMBINANT fusion proteins are essential tools in molecular studies across all organisms. Fusing fluorescent proteins to a protein of interest (POI) allows for direct visualization in situ. Fusing peptide epitopes, for which antibodies have been developed, to the POI is fundamental to many techniques, including Western blots, immunofluorescence, co-immunoprecipitation, chromatin immunoprecipitation, and purification (Arnau et al. 2006;Young et al. 2012;Bell et al. 2013).We describe an approach to add a protein or peptide epitope to a POI. One challenge to epitope tagging is choosing the location to attach the epitope to the POI. The standard design appends the epitope to the N-or C-terminus of the POI. However, in some instances, proteins cannot be tagged at the N or C terminus because a tag interferes with function, disrupting, for example, trafficking or post-translational modification. Tags can also interfere with protein folding or structure and disrupt protein-protein interactions.Our efforts to study the localization and function of Dcw1 in Saccharomyces cerevisiae have been hampered because we cannot introduce a tag at either terminus. Dcw1 and its paralog Dfg5 are important in cell-wall structure and integrity in S. cerevisiae and other fungi (Kitagaki et al. 2002(Kitagaki et al. , 2004Gonzalez et al. 2010;Maddi et al. 2012). Traditional methods of N-or C-terminally tagging Dcw1 are expected to fail due to interference with protein localization because Dcw1 localization at the cell membrane requires an N-terminal secretory signal sequence and a C-terminal signal sequence for addition of a glycosylphosp...

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Genome Evolution in Yeasts

Cited by 175 publications

References 98 publications

Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production

Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production

Research on the Relationship between Compensation Gaps of Executive-staff and Corporate Performance

Avoiding the Ends: Internal Epitope Tagging of Proteins Using Transposon Tn7

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