Production of hantavirus Puumala nucleocapsid protein in Saccharomyces cerevisiae for vaccine and diagnostics

Antoniukas, Linas; Grammel, Hartmut; Reichl, Udo

doi:10.1016/j.jbiotec.2005.12.028

Cited by 17 publications

(17 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Yet with all of these purported advantages, heterologous protein expression in yeast, in many instances, is far from optimal, with yields as low as micrograms per liter (11a, 35). However, yeast protein production capacity in general terms is much higher, given the ability to secrete certain proteins at levels approaching a gram per liter (1,24).…”

mentioning

confidence: 99%

A Novel High-Throughput Screen Reveals Yeast Genes That Increase Secretion of Heterologous Proteins

Wentz

Shusta

2007

Appl Environ Microbiol

109

View full text Add to dashboard Cite

The yeast Saccharomyces cerevisiae is an attractive host for the production of heterologous proteins. However, low-yield production of many proteins (from micrograms to milligrams/liter) leaves considerable room for optimization. By engineering the yeast cell via traceable genome-wide libraries, genes that can enhance protein expression level because of their roles in protein transcription, translation, folding, and trafficking processes can be readily identified. This report details a novel approach that combines yeast cDNA overexpression libraries with yeast surface display to allow the rapid flow cytometric screening of engineered yeast for gene products that improve the display of heterologous proteins. After optimization of the screening conditions, a genome-wide scan yielded five yeast gene products that promoted increased display levels of a single-chain T-cell receptor (scTCR). The display-enhancing genes included those coding for cell wall proteins (CCW12, CWP2, and SED1), a ribosomal subunit protein (RPP0), and an endoplasmic reticulum-resident protein (ERO1). Under the premise that yeast surface display levels could be used as a predictor of secretion efficiency, each display-enhancing gene product was tested for its ability to affect secretion levels of multiple scTCR and single-chain antibodies (scFv). All of the selected yeast gene products were shown to promote increased secretion of active protein (1.5-fold to 7.9-fold), with CCW12 and ERO1 being the most generalizable enhancers of scFv/scTCR secretion.

show abstract

mentioning

confidence: 99%

A Novel High-Throughput Screen Reveals Yeast Genes That Increase Secretion of Heterologous Proteins

Wentz

Shusta

2007

Appl Environ Microbiol

109

View full text Add to dashboard Cite

show abstract

“…Continuous culture maintains cells in stationary state (chemostat) with a constant supply of culture medium and cells; it is usually designed for producing a high volume of low value products associated to growth, typically primary metabolites and biomass (Bondarev et al 2003 ;Antoniukas et al 2006 ;Desai et al 2006 ).…”

Section: Culture Medium Selectionmentioning

confidence: 99%

In Vitro Plant Cultures as Biofactories

Álvarez

2014

Plant Biotechnology for Health

View full text Add to dashboard Cite

In vitro plant cell cultures are defi ned as a culture in aseptic conditions of any part of the plant body in a nutritive media. Cells, differentiated and undifferentiated (calli and suspended cells) tissues, and organs can be maintained in vitro .Some of the advantages of in vitro cultures are their development and maintenance in defi ned conditions (nutrients, light, humidity, temperature), independent from environmental infl uences (pests, diseases, extreme weather, geographical location, etc.), and manipulated by experienced operators in confi ned facilities.Since its development, in vitro plant cell cultures have been an essential tool for studying plant metabolism, and to micropropagate plant species, maintaining their genetic background. Also, they were helpful to analyze and optimize the production of plant secondary metabolites.Secondary metabolites productive processes are conducted, with a different degree of success, in undifferentiated cultures and in cultures of transformed organs. Several strategies were displayed in order to increase yields as manipulation of the plant growth regulator balance, addition of precursors, biotransformation, and immobilization.Also, as plant cells have a different behavior in liquid culture compared with microorganisms and mammal cells, bioreactor design was adapted to develop plant cell cultures.

show abstract

“…A fused galactose inducible GAL10-PYK promoter has recently been used for the production of high levels of hantavirus Puumala nucleocapsid (N) protein (316 mg/l) (Antoniukas et al 2006), and the HeV and NiV nucleocapsid proteins from Hendra and Nipah viruses, has been productively expressed (18-20 mg/l) in S. cerevisiae under the control of the GAL7 promoter (Juozapaitis et al 2007). A GAL1 promoter has also been used to drive the expression of a heterologous dextranase gene in S. cerevisiae, in order to develop a novel toxicity bioassay (Li et al 2006).…”

Section: Controlled Expression Using Endogenous Promotersmentioning

confidence: 99%

Systems for applied gene control in Saccharomyces cerevisiae

et al. 2008

View full text Add to dashboard Cite

Saccharomyces cerevisiae is frequently used in biotechnology, including fermentative processes in food production, heterologous protein production and high throughput developments for biomedicine. Accurate expression of selected genes is essential for all these areas. Systems that can be regulated are particularly useful because they allow controlling the timing and levels of gene expression. We examine here new expression systems that have been described, including improvements of classical ones and new strategies of artificial gene control that have been applied in functional genomics.

show abstract

Production of hantavirus Puumala nucleocapsid protein in Saccharomyces cerevisiae for vaccine and diagnostics

Cited by 17 publications

References 44 publications

A Novel High-Throughput Screen Reveals Yeast Genes That Increase Secretion of Heterologous Proteins

A Novel High-Throughput Screen Reveals Yeast Genes That Increase Secretion of Heterologous Proteins

In Vitro Plant Cultures as Biofactories

Systems for applied gene control in Saccharomyces cerevisiae

Contact Info

Product

Resources

About