Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1995-05-25
1998-03-10
Ketter, James
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
435 694, 435 711, 43525411, 4352552, 4353201, 536 231, 536 235, 536 2351, C12N 1500
Patent
active
057260389
DESCRIPTION:
BRIEF SUMMARY
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a of PCT/DK94/00281 filed Jul. 8, 1994, which is incorporated herein by reference.
FIELD OF INVENTION
The present invention relates to a DNA construct comprising the YAP3 signal peptide for secretion of a heterologous polypeptide, a yeast cell containing the DNA construct and a method of producing heterologous polypeptides in yeast from the DNA construct.
BACKGROUND OF THE INVENTION
Yeast organisms produce a number of proteins which are synthesized intracellularly, but which have a function outside the cell. Such extracellular proteins are referred to as secreted proteins. These secreted proteins are expressed initially inside the cell in a precursor or a pre-protein form containing a presequence ensuring effective direction of the expressed product across the membrane of the endoplasmic reticulum (ER). The presequence, normally named a signal peptide, is cleaved off from the rest of the protein during translocation. Once entered in the secretory pathway, the protein is transported to the Golgi apparatus. From the Golgi the protein can follow different routes that lead to compartments such as the cell vacuole or the cell membrane, or it can be routed out of the cell to be secreted to the external medium (Pfeffer, S. R. and Rothman, J. E. Ann. Rev. Biochem. 56 (1987), 829-852).
Several approaches have been suggested for the expression and secretion in yeast of proteins heterologous to yeast. European published patent application No. 88 632 describes a process by which proteins heterologous to yeast are expressed, processed and secreted by transforming a yeast organism with an expression vehicle harbouring DNA encoding the desired protein and a signal peptide, preparing a culture of the transformed organism, growing the culture and recovering the protein from the culture medium. The signal peptide may be the signal peptide of the desired protein itself, a heterologous signal peptide or a hybrid of native and heterologous signal peptide.
A problem encountered with the use of signal peptides heterologous to yeast might be that the heterologous signal peptide does not ensure efficient translocation and/or cleavage after the signal peptide.
The S. cerevisiae MF.alpha.1 (.alpha.-factor) is synthesized as a prepro form of 165 amino acids comprising signal-or prepeptide of 19 amino acids followed by a "leader" or propeptide of 64 amino aicds, encompassing three N-linked glycosylation sites followed by (LysArg(Asp/Glu, Ala).sub.2-3 .alpha.-factor).sub.4 (Kurjan, J. and Herskowitz, I. Cell 30 (1982), 933-943). The signal-leader part of the preproMF.alpha.1 has been widely employed to obtain synthesis and secretion of heterologous proteins in S. cerivisiae.
Use of signal/leader peptides homologous to yeast is known from i.a. U.S. Pat. No. 4,546,082, European published patent applications Nos. 116 201, 123 294, 123 544, 163 529, and 123 289 and DK patent application No. 3614/83.
In EP 123 289 utilization of the S. cerevisiae a-factor precursor is described whereas WO 84/01153 indicates utilization of the Saccharomyces cerevisiae invertase signal peptide and DK 3614/83 utilization of the Saccharomyces cerevisiae PH05 signal peptide for secretion of foreign proteins.
U.S. Pat. No. 4,546,082, EP 16 201, 123 294,123 544, and 163 529 describe processes by which the .alpha.-factor signal-leader from Saccharomyces cerevisiae (MF.alpha.1 or MF.alpha.2) is utilized in the secretion process of expressed heterologous proteins in yeast. By fusing a DNA sequence encoding the S. cerevisiea MF.alpha.1 signal/leader sequence at the 5' end of the gene for the desired protein secretion and processing of the desired protein was demonstrated.
A number of secreted proteins are routed so as to be exposed to a proteolytic processing system which can cleave the peptide bond at the carboxy end of two consecutive basic amino acids. This enzymatic activity is in S. cerevisiae encoded by the KEX 2 gene (Julius, D. A. et al., Cell 37 (1984b), 1075). Processing of the product by the
REFERENCES:
patent: 5037743 (1991-08-01), Welch et al.
patent: 5217891 (1993-06-01), Brake et al.
patent: 5538863 (1996-07-01), Price
Egel-Mitani, M. 1990 Yeast 6:127-137.
J. Cellular Biochem., vol. 12, (1988) Suppl. O. Part B.
Christiansen Lars
Petersen Jens Gunner Litske
Gregg, Esq. Valeta
Ketter James
Novo Nordisk A S
Yucel Irem
Zelson Esq. Steve T.
LandOfFree
DNA construct encoding the YAP3 signal peptide does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with DNA construct encoding the YAP3 signal peptide, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and DNA construct encoding the YAP3 signal peptide will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-138476