Cloning and expression of a biologically active Anisakis simplex allergen Ani s 1 in the yeast Pichia pastoris

2007 ◽  
Vol 154 (1) ◽  
pp. 115-118 ◽  
Author(s):  
Rosa Rodriguez-Perez ◽  
Maria Luisa Caballero ◽  
Miguel Gonzalez-Munoz ◽  
Ana Rodriguez-Mahillo ◽  
Ignacio Moneo
Keyword(s):  
Pichia Pastoris ◽  
Anisakis Simplex ◽  
Biologically Active ◽  
Cloning And Expression ◽  
Yeast Pichia Pastoris

The bovine IFN-ω1 is biologically active and secreted at high levels in the yeast Pichia pastoris

1998 ◽  
Vol 60 (1-2) ◽  
pp. 3-14 ◽  
Author(s):  
Manuel Rodrı́guez ◽  
Vladimir Martı́nez ◽  
Katy Alazo ◽  
Marisela Suárez ◽  
Miguel Redondo ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Biologically Active ◽  
Yeast Pichia Pastoris

scholarly journals Production of biologically active tethered ovine FSHbetaalpha by the methylotrophic yeast Pichia pastoris

2003 ◽  
Vol 30 (2) ◽  
pp. 213-225 ◽  
Author(s):  
AE Fidler ◽  
JS Lin ◽  
S Lun ◽  
W Ng Chie ◽  
A Western ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Fusion Protein ◽  
Fusion Gene ◽  
Methylotrophic Yeast ◽  
Biologically Active ◽  
Single Chain ◽  
Glycoprotein Hormone ◽  
Yeast Pichia Pastoris ◽  
Methylotrophic Yeast Pichia Pastoris

The pituitary-derived glycoprotein hormone FSH plays a central role in controlling vertebrate gonadal function. In female mammals the maturation of ovarian follicles is critically dependent upon stimulation by FSH. Moreover, injection of exogenous FSH is used extensively to stimulate increased numbers of follicles to ovulate. Structurally FSH is a heterodimeric glycoprotein composed of two non-covalently associated polypeptide subunits. The tertiary structures of both the alpha- and beta-subunits are constrained by intramolecular disulphide bonds and are post-translationally modified with two N-linked carbohydrate moieties, the structure of which appears to modulate in vivo biological activity. Here we report the expression of ovine FSH (oFSH) as a biologically active single-chain polypeptide using the methylotrophic yeast Pichia pastoris. Sequences encoding the mature oFSH alpha- and beta-proteins were fused to form a gene encoding a fusion protein with the C-terminus of the beta-chain joined to the N-terminus of the alpha-chain, with the chains separated by a two amino acid linker sequence. This fusion gene was itself fused to two alternative Pichia leader sequences (mating factor alpha and acid phosphatase) and transformed into the Pichia strains GS115 and SMD1168. The recombinant fusion protein (oFSHbetaalpha) was expressed at approximately 0.1 microg/ml in 'shake-flask' cultures. The Pichia-expressed tethered protein was biologically active in an in vitro bioassay, had a molecular mass of 28 kDa, as determined by SDS-PAGE, and bound the bovine FSH receptor with a binding profile similar to that of native oFSH.

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