Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Patent
1993-03-25
1996-03-19
Schwartz, Richard A.
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
435 691, 43525411, 4352542, 43525421, 43525422, 43525423, C12N 1511, C12N 119
Patent
active
055004830
ABSTRACT:
Two novel genomic clones, ZZA1 and ZZA2, were isolated which encode for Pichia pastoris alcohol oxidase isozymes. The 5' non-coding region of ZZA1 contains common structural features involved in the transcription and translation of eukaryotic genes. Comparison of the nucleotide sequences of the ZZA1 and AOX15' noncoding regions showed that they are 66% similar to each other.
The rice .alpha.-amylase gene OS103 was placed under the transcriptional control of the ZZA1 promoter. The nucleotide sequences of ZZA1 and other methanol-regulated promoters were analyzed. A highly conserved sequence (TTGNNNGCTTCCAANNNNNTGGT) (SEQ ID NO: 2) was found in the 5' flanking region. A yeast strain containing the ZZA1-OS103 fusion and secreting biologically active .alpha.-amylase into the culture media while converting starch to ethanol was produced. The ZZA1 and ZZA2 regulatory sequences may be used to contol the expression of other heterologous proteins in multiple yeast species.
Methods of purifying proteins that are regulators of alcohol oxidase expression (referred to as AOER proteins) and methods of isolating these proteins are also provided.
REFERENCES:
patent: 4808537 (1989-02-01), Stroman et al.
Ellis, et al., "Isolation of alcohol Oxidase and Two Other Methanol Regulatable Genes from the Yeast Pichia pastoris", Mol. Cell Biol. 5:1111-1121 (1985).
Tschopp, et al., "Expression of the 1acZ gene from two methanol-regulated promoters in Pichia pastoris", Nucleic Acids Research 15: 3859-3876 (1987).
Cregg, et al., "High-Level Expression and Efficient Assembly of Hepatitis B Surface Antigen in The Methylotrophic Yeast, Pichia Pastoris", Bio/Technology 5: 479-485 (1987).
Tschopp, et al., "High-level Secretion of Glycosylated Invertase In The Methylotrophic Yeast, Pichia Pastoris", Bio/Technology 5: 1305-1308 (1987).
Sreekrishna, et al., "High-Level Expression, Purification, and Characterization of Recombinant Human Tumor necrosis Factor Synthesized in the Methylotrophic Yeast Pichia Pastoris", Biochemistry 28: 4117-4125 (1989).
Clare, et al., "High-Level Expression of Tetanus Toxin Fragment C in Pichia Pastoris Strains Containing Multiple Tandem Integrations of the Gene", Bio/Technology 9: 455-460 (1991).
Sakai and Tani, "Cloning and sequencing of the alcohol oxidas-encoding gene (AOD1) from the formaldehyde-producing asporogenous methylotrophic yeast, Candida boidinii S2", Gene 114: 67-73 (1992).
Ledeboer, et al., "Molecular cloning and characterization of a gene coding for methanol oxidase in Hansenula polymorpha", Nucleic Acids Research 13: 3063-3082 (1985).
Koutz, et al., "Structural Comparison of the Pichia pastoris Alcohol Oxidase Gene", Yeast 5: 167-177 (1989).
Buchman, et al., "Two DNA-Binding Factors Recognize Specific Sequences at Silencers, Upstream Activating Sequences, Autonomously Replicating Sequences, and Telomeres in in Saccharomyces cerevisiae", Mol. Cell. Biol. 8: 210-225 (1988).
Brindle, et al., "Multiple Factors Bind the Upstream Activation Sites of the Yeast Enolase GFenes ENO1 and ENO2: ABFI Protein, like Repressor Activator Protein RAP1, Binds cis-Acting Sequences Which Modulate Repression or Activation of Transcription", Mol. Cell. Biol. 10: 4872-4885 (1990).
Huie, et al., "Characterization of the DNA-Binding Activity of GCR1: In vivo Evidence for Two GCR1-Binding Sites in the Upstream Activiating Sequence of TPI of Saccharomyces cerevisae", Mol. Cell. Biol. 12: 2690-2700 (1992).
Laimens, et al., Enhances and Eukaryotic Gene Expression: Current Communications in Molecular Biology, Gluzman and Shenk, eds. Cold Spring Harbor Press, New York (1983).
Cohen, et al., "Transcription of the Constitutively Expressed Yeast Enolase Gene ENO1 Is Mediated by Positive and Negative cis-Acting Regulatory Sequences", Mol. Cell. Biol. 7:2753-2761 (1987).
Cregg, et al., "Functional Characterization of the Two Alcohol Oxidase Genes from the Yeast Pichia pastoris", Mol. Cell. Biol. 9: 1316-1323 (1989).
Olesen, et al., "Yeast HAP2 and HAP3 Activators Both Bind to the CYC1 Upstream Activation Site, UAS2, in an Interdependent Manner", Cell 51: 953-961 (1987).
Hahn and Guarente, "Yeast HAP2 and HAP3: Transcriptional Activators in a Heteromeric Complex", Science 240: 317-321 (1988).
Kumagai et. al. 1990, Gene 94: 209-216.
Guarente 1987 Ann. Rev. Genet, 21 425-452.
Kumagai Monto H.
Sverlow Genadie G.
Biosource Technologies, Inc.
Halluin Albert P.
Schwartz Richard A.
Vogel Nancy T.
LandOfFree
Pichia pastoris alcohol oxidase ZZA1 regulatory region for heter does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Pichia pastoris alcohol oxidase ZZA1 regulatory region for heter, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Pichia pastoris alcohol oxidase ZZA1 regulatory region for heter will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1959831