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Leader sequence for yeast

A sequence and leader peptide technology, applied in the field of leader sequences for yeast, can solve the problems of the limited number of secretory sequences of yeast and the lack of extensive characterization

Pending Publication Date: 2021-06-22
BASF AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Since most yeast do not secrete large numbers of endogenous proteins and their extracellular proteomes have not been extensively characterized so far, the number of secretion sequences available for yeast is limited

Method used

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  • Leader sequence for yeast
  • Leader sequence for yeast
  • Leader sequence for yeast

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[0129] 1. General method for expression in P. phaaffiae (Pichia pastoris)

[0130] The leader sequence was cloned into the pPICz backbone (Thermo Fischer) upstream of the gene of interest (eg lipase, amylase or xylanase). The expression of the gene of interest is regulated by the methanol-inducible AOX1 promoter present in the pPICz backbone or the methanol-free constitutive promoter according to SEQ ID No: 11 cloned into the pPICz backbone to replace the AOX1 promoter. The expression vector was transformed into P. phaaffiae strain X-33 and selected for transformation by zeocin selection as described in: User Manual for pPICZ A, B, and C A, B and C), Revised July 7, 2010, Manual Part No. 25-0148. Cultures in microtiter plates in YPD medium (1% yeast extract, 2% peptone, 2% glucose in sterile water) with plasmids comprising the methanol-free constitutive promoter according to SEQ ID No: 11 Single colonies of transformed strains. In microtiter plates in BMMY medium (2% pepton...

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Abstract

The present invention relates to a leader peptide which promotes the secretion of recombinant proteins and a nucleic acid sequence encoding the leader peptide as well as expression cassettes, vectors and host cells comprising this leader sequence. Also disclosed is a method for producing a protein using this leader peptide.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of priority to US Application No. 62 / 769,242, filed November 19, 2018, the contents of which are incorporated herein by reference in their entirety. [0003] sequence listing [0004] The present application contains the content according to "Standard for the Presentation of Nucleotide and Amino Acid Sequence Listings in International Patent Applications Under the Patent Cooperation Treaty (PCT)" "ST.25 Nucleotide and amino acid sequence listing in computer readable form (CRF) as an ASC II text (.txt) file. The Sequence Listing is identified below and is incorporated by reference in its entirety into this specification of the present application for all purposes. [0005] file name creation date size (bytes) 180263US01_SequenceListing.txt 11 / 16 / 2018 32KB (32,847 bytes) technical field [0006] The present invention relates to leader peptides that facilit...

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Application Information

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IPC IPC(8): C11D3/386C12N9/20C12R1/84
CPCC12N9/20C12N2800/102C12N2810/70C12P21/02C12N15/815C07K14/395C07K14/39C07K2319/02C12N15/625C12R2001/84C12N1/165C12N9/242C12N9/2482C12Y301/01C12Y302/01001C12Y302/01008
Inventor 谭旭秋钟静萍M·A·斯克兰顿
Owner BASF AG