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Construction and application of the high sensitivity expression system of copper-resistant Saccharomyces cerevisiae(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2014 03
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Construction and application of the high sensitivity expression system of copper-resistant Saccharomyces cerevisiae
SHENG Guanyi ZHUGE Bin ZONG Hong LU Xinyao FANG Huiying SONG Jian ZHUGE Jian
1Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China 2School of Biotechnology, Research Centre of Industrial Microbiology, Jiangnan University, Wuxi 214122, China 3School of Chemical and Material Engineering, Jiangnan University, Wuxi 214122, China
Saccharomyces cerevisiae PCR-mediated Technique cup1 copper-resistance gfp
Q939.97 : Q786

This study was aimed to construct a high sensitivity expression system of copper-resistant Saccharomyces cerevisiae. With the PCR-mediated technique which allows single-step deletion of chromosomal gene to knock out the cup1 gene of S. cerevisiae W303-1A, the lowest inhibition concentration of Cu2+ was determined by gradient dilution. The construction of the expression system pYX212M used pYX212 as the basic frame, cup1 as the selective gene and the S. cerevisiae W303-1A cup1D as the host. The pYX212MGFP was constructed, transforming S. cerevisiae W303-1A cup1D to observe whether the strain displayed green fluorescence. A copper-sensitive S. cerevisiae W303-1A cup1D was obtained and its lowest inhibition concentration of Cu2+ was 1.2 mmol/L compared with 0.08 mmol/L of S. cerevisiae W303-1A. The expression system pYX212M was achieved. The green fluorescence was seen under the microscope by transforming pYX212MGFP into the S. cerevisiae W303-1A cup1D, demonstrating the successful expression of gfp. The results suggested that the high sensitivity expression system of copper-resistant S. cerevisiae can be constructed. It not only enriches the expression system of S. cerevisiae, but also provides guidance in the research of the copper resistance mechanisms and the bioremediation function of the microorganism.


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