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Construction of Response Vectors in Yeast for Environmental Estrogen Bioassay

Shu Zeng, Yong Diao, Qing Song Li, Hui Yang

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Abstract

The environmental estrogen can be measured in modified yeast cells by transcription activation of reporter genes. To establish such yeast estrogen system, the full-length human estrogen receptor (hER) cDNA and estrogen responsive elements (EREs) linked to the upstream of CYC1 promoter were cloned into the yeast expression vector via pPIC9 vector respectively to construct the response vectors named as pER-PIC9 (E-9) and pERE2-CYC-PIC9K-EGFP (EC-9-E), which were transformed into Pichia pastoris at the same time for detecting environmental estrogens based on yeast Enhanced Green Fluorescence Protein (yEGFP) as a reporter protein. In the present study, we developed a rapid yeast estrogen bioassays by creating stably transfected strains with EGFP as a measurable reporter protein.

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What this paper is about

The environmental estrogen can be measured in modified yeast cells by transcription activation of reporter genes. To establish such yeast estrogen system, the full-length human estrogen receptor (hER) cDNA and estrogen responsive elements (EREs) linked to the upstream of CYC1 promoter were cloned into the yeast expression vector via pPIC9 vector respectively to construct the response vectors named as pER-PIC9 (E-9) and pERE2-CYC-PIC9K-EGFP (EC-9-E), which were transformed into Pichia pastoris at the same time for detecting environmental estrogens based on yeast Enhanced Green Fluorescence Protein (yEGFP) as a reporter protein. In the present study, we developed a rapid yeast estrogen bioassays by creating stably transfected strains with EGFP as a measurable reporter protein.

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Available abstract

The environmental estrogen can be measured in modified yeast cells by transcription activation of reporter genes. To establish such yeast estrogen system, the full-length human estrogen receptor (hER) cDNA and estrogen responsive elements (EREs) linked to the upstream of CYC1 promoter were cloned into the yeast expression vector via pPIC9 vector respectively to construct the response vectors named as pER-PIC9 (E-9) and pERE2-CYC-PIC9K-EGFP (EC-9-E), which were transformed into Pichia pastoris at the same time for detecting environmental estrogens based on yeast Enhanced Green Fluorescence Protein (yEGFP) as a reporter protein. In the present study, we developed a rapid yeast estrogen bioassays by creating stably transfected strains with EGFP as a measurable reporter protein.

Key concepts: Yeast, Reporter gene, Pichia pastoris, Expression vector, Estrogen receptor, Estrogen, Green fluorescent protein, Biology

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