Use of the cysteine-repressible HpMET3 promoter as a novel tool to regulate gene expression in Hansenula polymorpha.
Biotechnol Lett
; 37(11): 2237-45, 2015 Nov.
Article
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| MEDLINE
| ID: mdl-26169200
ABSTRACT
OBJECTIVES:
The promoter of HpMET3, encoding an ATP sulfurylase, was evaluated for its potential as a repressible promoter to downregulate the expression of target genes in the thermotolerant, methylotrophic yeast Hansenula polymorpha.RESULTS:
The expression of lacZ under the control of the 0.6 kb HpMET3 promoter was efficiently downregulated by cysteine, but not by methionine or sulfate. The HpMET3 promoter was used to generate a conditional mutant of the HpPMT2 gene encoding an O-mannosyltransferase, which is involved in post-translational protein modification. The addition of 0.5 mM cysteine adversely affected the growth of the conditional HpMET3(p)-Hppmt2 mutant strain by downregulating transcription of HpPMT2 to approx. 40 % of the normal levels, indicating that the HpPMT2 gene is essential for cell viability. However, the HpMET3 promoter was neither induced nor repressed in the heterologous host Saccharomyces cerevisiae.CONCLUSION:
Our results reveal that the cysteine-repressible HpMET3 promoter is a useful tool that downregulates the expression of various genes in H. polymorpha.Palabras clave
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Banco de datos:
MEDLINE
Asunto principal:
Pichia
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Ingeniería Genética
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Regulación Fúngica de la Expresión Génica
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Regiones Promotoras Genéticas
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Cisteína
Idioma:
En
Revista:
Biotechnol Lett
Año:
2015
Tipo del documento:
Article