(YY020) YLEX Yeast Expression Kit

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2 570,00 € 2570.0 EUR 2 570,00 € hors TVA

2 570,00 € hors TVA

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    Garantie satisfait ou remboursé de 30 jours
    Expédition : 2-3 jours ouvrables

    General background of Yarrowia lipolytica:

    Yarrowia lipolytica is a species of non-conventional and GRAS (generally regarded as

    safe) yeast widely utilized in industrial applications such as organic acid and protein

    production. As unicellular organism, it has the advantages of E. coli and Saccharomyces

    cerevisiae in ease of manipulation and growth capacity. But, it also functions as a higher

    eukaryotic organization able to perform post-translational processing of complex

    proteins. As compared to S. cerevisiae, Y. lipolytica has certain advantages, such as a

    mainly co-translational secretion pathway (like in mammalian cells), higher secretion

    capacity and product yield, less hyperglycosylation on products, and simplicities in

    scaling-up production. These features make Y. lipolytica very useful as a protein

    expression system. Furthermore, the whole genome of Yarrowia lipolytica has been

    sequenced; please check http://cbi.labri.fr/Genolevures/elt/YALI for details.

    Product Description:

    YLEX Expression Kit, based on INRA INAPG licensed patent*, provides an easy approach for cloning and expressing a gene of interest in the yeast Yarrowia lipolytica. Using this kit, heterologous protein may be expressed intracellularly or secreted from the cell into  medium  by  selecting  respectively  the  supplied  expression  vector  pYLEX1  or pYLSC1. 

    Using  YLEX Expression Kit, heterologous protein expression is driven by a strong hybrid promoter (hp4d) carrying four tandem copies of an upstream activator sequence (UAS1B) from pXPR2 and a minimal pLEU2 fragment. To achieve expression in yeast,  pYLEX1  containing  a  cloned  gene  of  interest  is  linearized  by  a  selected restriction  enzyme  to  produce  an  expression  cassette  that  can  integrate  with  high efficiency  into  the  Y. lipolytica genome by homologous recombination with an integrated pBR platform. A leucine gene (LEU2) in pYLEX1 provides for selection of yeast containing an integrated expression cassette by allowing their growth on leucine-free minimal medium. The integrated vector is particularly stable, and targeted monocopy integration allows a direct comparison of the properties of the transformants, which are comparable in terms of locus and copy number.

    To achieve secretion of protein from yeast cells, the gene of interest is cloned into pYLSC1,  downstream  from  the  XPR2 pre region (secretion signal from XPR2 gene),  resulting in expression of a secretion signal fusion protein. The XPR2 pre region directs the fusion protein to be eciently transported through the yeast secretory pathway. The secretion signal fusion protein undergoes sequential processing by signal peptidase and protease in the endoplasmic reticulum and Golgi complex respectively, resulting in the secretion of the native form of the protein of interest into the culture medium.