pYLSC1解脂耶氏酵母表达载体质粒 BioVector NTCC质粒载体菌种细胞基因保藏中心

pYLSC1解脂耶氏酵母表达载体质粒

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, high level of heterologous protein may be expressed intracellularly or be secreted from the cell into the medium by selecting the supplied expression vector pYLEX1 or pYLSC1.


* INRA (Institut National de la Recherche Agronomique) and INAPG (Institut National Agronomique Paris-Grignon)

Features
Safe: Y. lipolytica was classified as GRAS (Generally Regarded As Safe) by the US FDA (Food and Drug Administration)
Simple: a simple tool for expressing heterologous protein
Easy manipulation: like E. coli and S. cerevisiae
Stable: strong stability in vectors and constructed plasmids
Reliable: vectors integrated at the same site in genome
Flexible: both expression and secretion vector provided (proteins may be expressed intracellularly or be secreted from the cell into medium)
High growth ability: high secretion capacity & high product yield
Less protein degraded: no extracellular protease synthesized by a special protease-deficient Yarrowia strain
Mass production: industrial mass production of recombinant proteins
Less hyperglycosylation:able to perform post-translational processing of complex proteins, unlike S. cerevisiae
Applications
Heterologous protein expression, either intracellular or extracellular depends on selected vector in GRAS yeast.
Yarrowia Vectors
Two vectors (pYLEX1 and pYLSC1) are included in this kit, and they can be used for either intracellular expression or secretion of proteins of interest in Y. lipolytica. Generally speaking, if the target protein is cytosolic and non-glycosylated, the pYLEX1 vector is a better choice. If the protein of your interest is normally glycosylated or secreted, you may want to choose the pYLSC1 vector.

The pYLEX1 expression vector (7259 bp) contains the strong hybrid promoter (hp4d) carrying four tandem copies of upstream activator sequences (UAS1B) fragment from pXPR2 and a minimal pLEU2 fragment. The multiple cloning sites and the XPR2 transcription terminator lie immediately downstream of 3′ site of hp4d promoter, followed by a leucine selection marker gene (LEU2). The vector can be linearized by digestion with NotI to create a linear DNA fragment capable of inserting into the Y. lipolytica genome.

The pYLSC1 secretion vector (7205 bp) contains the hybrid promoter (hp4d) and a secretion signal (XPR2 pre region). The multiple cloning site and the pXPR2 transcription terminator lie immediately downstream of 3′ site of XPR2 pre region, followed by a leucine selection marker gene (LEU2). The vector can be linearized by digestion with NotI to create a linear DNA fragment capable of inserting into the Y. lipolytica genome.
Yeast Strain
The strain Po1g of Yarrowia lipolytica is a derivative of the wild-type strain W29 (ATCC 20460) by a series of genetic modifications. Briefly, the original URA3 gene in the W29 strain was disrupted with the SUC2 gene from Saccharomyces cerevisiae, followed by the introduction of a deletion in the LEU2 gene. Furthermore, the deletion of the XPR2 and AXP genes ensures that Po1g is unable to produce any extracellular protease. In order to allow easy integration of pBR-based expression/secretion vectors, a pBR322 docking platform was integrated at the URA3 locus.

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