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Amuel, Carsten,Gellissen, Gerd,Cor,Suckow, Manfred The Korean Society for Biotechnology and Bioengine 2000 Biotechnology and Bioprocess Engineering Vol.5 No.4
The strength and regulatory characteristics of the heat-inducible HSA1, HSA2 and TPS1 promoters were compared with those of the well-established, carbon source-regulated FMD promoter in a Hansenula polymorpha-based host system in vivo. In addition, the Saccharomyces cerevisiae-derived ADH1 promoter was analysed. While ADH1 promoter showed to be of poor activity in the foreign host, the strength of the heat shock TPS1 promoter was found to exceed that of the FMD promoter, which at present is considered to be the strongest promoter for driving heterologous gene expression in H. polymorpha.
Park, Jeong-Nam,Sohn, Min Jeong,Oh, Doo-Byoung,Kwon, Ohsuk,Rhee, Sang Ki,Hur, Cheol-Goo,Lee, Sang Yup,Gellissen, Gerd,Kang, Hyun Ah American Society for Microbiology 2007 Applied and environmental microbiology Vol.73 No.19
<B>ABSTRACT</B><P>The genomewide gene expression profiling of the methylotrophic yeast <I>Hansenula polymorpha</I> exposed to cadmium (Cd) allowed us to identify novel genes responsive to Cd treatment. To select genes whose promoters can be useful for construction of a cellular Cd biosensor, we further analyzed a set of <I>H. polymorpha</I> genes that exhibited >6-fold induction upon treatment with 300 μM Cd for 2 h. The putative promoters, about 1,000-bp upstream fragments, of these genes were fused with the yeast-enhanced green fluorescence protein (GFP) gene. The resultant reporter cassettes were introduced into <I>H. polymorpha</I> to evaluate promoter strength and specificity. The promoter derived from the <I>H. polymorpha SEO1</I> gene (Hp<I>SEO1</I>) was shown to drive most strongly the expression of GFP upon Cd treatment among the tested promoters. The Cd-inducible activity was retained in the 500-bp deletion fragment of the Hp<I>SEO1</I> promoter but was abolished in the further truncated 250-bp fragment. The 500-bp Hp<I>SEO1</I> promoter directed specific expression of GFP upon exposure to Cd in a dose-dependent manner, with Cd detection ranging from 1 to 900 μM. Comparative analysis of the <I>Saccharomyces cerevisiae SEO1</I> (Sc<I>SEO1</I>) promoter revealed that the Sc<I>SEO1</I> promoter has a broader specificity for heavy metals and is responsive to arsenic and mercury in addition to Cd. Our data demonstrate the potential use of the Hp<I>SEO1</I> promoter as a bioelement in whole-cell biosensors to monitor heavy metal contamination, particularly Cd.</P>