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.
journal_name
Biotechnol Lettjournal_title
Biotechnology lettersauthors
Yoo SJ,Chung SY,Lee DJ,Kim H,Cheon SA,Kang HAdoi
10.1007/s10529-015-1902-5subject
Has Abstractpub_date
2015-11-01 00:00:00pages
2237-45issue
11eissn
0141-5492issn
1573-6776pii
10.1007/s10529-015-1902-5journal_volume
37pub_type
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