Difference between revisions of "Part:BBa K2924039"
 
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Methanol is processed in P. pastoris by an alcohol oxidases (AOX) converting it to formaldehyde, which are coded by AOX genes (Tschopp et al., 1987). The AOX1 promotor is highly induced by methanol, repressed in the presence of other carbon sources like glucose, and therefore genes under the control of the promotor are highly expressed in the presence of methanol and can be easily regulated (Tschopp et al., 1987; Daly and Hearn, 2005). Methylotrophic promotors are among the strongest and strictly regulated yeast promotors, which makes them suitable for controlled strong expression of target proteins after methanol induction (Demain and Vaishnav, 2011). The AOX1 promotor can produce a high yield of recombinant proteins up to 30 % of the total mass of cell or up to 22 g/L intercellular protein in P. pastoris (Romanos et al., 1992; Vogl et al., 2013).
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Methanol is processed in <I>P. pastoris</I> by alcohol oxidases (AOX) converting it to formaldehyde, which are coded by <i>AOX</i> genes<sup>1</sup>. The <i>AOX1</i> promotor is highly induced by methanol, repressed in the presence of other carbon sources like glucose, and therefore genes under the control of the promotor are highly expressed in the presence of methanol and can be easily regulated<sup>2,3</sup>. Methylotrophic promotors are among the strongest and strictly regulated yeast promotors, which makes them suitable for controlled strong expression of target proteins after methanol induction<sup>4</sup>. The <i>AOX1</i> promotor can produce a high yield of recombinant proteins up to 30 % of the total mass of cell or up to 22 g/L intercellular protein in <I>P. pastoris</I><sup>5</sup>.
  
 
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===References===
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===Functional Parameters===
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<partinfo>BBa_K2924039 parameters</partinfo>
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1: Ebina Y, Edery M, Ellis L, Standring D, Beaudoin J, Roth RA, Rutter WJ (1985) Expression of a functional human insulin receptor from a cloned cDNA in Chinese hamster ovary cells. Proc Natl Acad Sci U S A 82: 8014–8018
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2: Demain AL, Vaishnav P (2009) Production of recombinant proteins by microbes and higher organisms. Biotechnol Adv 27: 297–306
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3: Terpe K (2006) Overview of bacterial expression systems for heterologous protein production: From molecular and biochemical fundamentals to commercial systems. Appl Microbiol Biotechnol 72: 211–222
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4: Tran A-M, Nguyen T-T, Nguyen C-T, Huynh-Thi X-M, Nguyen C-T, Trinh M-T, Tran L-T, Cartwright SP, Bill RM, Tran-Van H (2017) Pichia pastoris versus Saccharomyces cerevisiae: a case study on the recombinant production of human granulocyte- macrophage colony-stimulating factor. BMC Res Notes 10: 148
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5: Byrne B (2015) Pichia pastoris as an expression host for membrane protein structural biology. Curr Opin Struct Biol 32: 9–17 Lin-Ceregh

Latest revision as of 16:33, 21 October 2019


AOX1 methanol-inducible promoter

Methanol is processed in P. pastoris by alcohol oxidases (AOX) converting it to formaldehyde, which are coded by AOX genes1. The AOX1 promotor is highly induced by methanol, repressed in the presence of other carbon sources like glucose, and therefore genes under the control of the promotor are highly expressed in the presence of methanol and can be easily regulated2,3. Methylotrophic promotors are among the strongest and strictly regulated yeast promotors, which makes them suitable for controlled strong expression of target proteins after methanol induction4. The AOX1 promotor can produce a high yield of recombinant proteins up to 30 % of the total mass of cell or up to 22 g/L intercellular protein in P. pastoris5.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

1: Ebina Y, Edery M, Ellis L, Standring D, Beaudoin J, Roth RA, Rutter WJ (1985) Expression of a functional human insulin receptor from a cloned cDNA in Chinese hamster ovary cells. Proc Natl Acad Sci U S A 82: 8014–8018

2: Demain AL, Vaishnav P (2009) Production of recombinant proteins by microbes and higher organisms. Biotechnol Adv 27: 297–306

3: Terpe K (2006) Overview of bacterial expression systems for heterologous protein production: From molecular and biochemical fundamentals to commercial systems. Appl Microbiol Biotechnol 72: 211–222

4: Tran A-M, Nguyen T-T, Nguyen C-T, Huynh-Thi X-M, Nguyen C-T, Trinh M-T, Tran L-T, Cartwright SP, Bill RM, Tran-Van H (2017) Pichia pastoris versus Saccharomyces cerevisiae: a case study on the recombinant production of human granulocyte- macrophage colony-stimulating factor. BMC Res Notes 10: 148

5: Byrne B (2015) Pichia pastoris as an expression host for membrane protein structural biology. Curr Opin Struct Biol 32: 9–17 Lin-Ceregh