A novel constructed SPT15 mutagenesis library of Saccharomyces cerevisiae by using gTME technique for enhanced ethanol production
Abstract During the last few years, the global transcription machinery engineering (gTME) technique has gained more attention as an effective approach for the construction of novel mutants. Genetic strategies (molecular biology methods) were utilized to get mutational for both genes (SPT15 and TAF23...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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SpringerOpen
2017-06-01
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| Series: | AMB Express |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13568-017-0400-7 |
| _version_ | 1811206752052445184 |
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| author | Ashraf A. M. M. El-Rotail Liang Zhang Youran Li Shuang Ping Liu Gui Yang Shi |
| author_facet | Ashraf A. M. M. El-Rotail Liang Zhang Youran Li Shuang Ping Liu Gui Yang Shi |
| author_sort | Ashraf A. M. M. El-Rotail |
| collection | DOAJ |
| description | Abstract During the last few years, the global transcription machinery engineering (gTME) technique has gained more attention as an effective approach for the construction of novel mutants. Genetic strategies (molecular biology methods) were utilized to get mutational for both genes (SPT15 and TAF23) basically existed in the Saccharomyces cerevisiae genome via screening the gTME approach in order to obtain a new mutant S. cerevisiae diploid strain. The vector pYX212 was utilized to transform these genes into the control diploid strain S. cerevisiae through the process of mating between haploids control strains S. cerevisiae (MAT-a [CICC 1374]) and (MAT-α [CICC 31144]), by using the oligonucleotide primers SPT15-EcoRI-FW/SPT15-SalI-RV and TAF23-SalI-FW/TAF23-NheI-RV, respectively. The resultant mutants were examined for a series of stability tests. This study showed how strong the effect of using strategic gTME with the importance of the modification we conducted in Error Prone PCR protocol by increasing MnCl2 concentration instead of MgCl2. More than ninety mutants we obtained in this study were distinguished by a high level production of bio-ethanol as compared to the diploid control strain. |
| first_indexed | 2024-04-12T03:53:25Z |
| format | Article |
| id | doaj.art-f0a6ffdaff8f44529334e4d42c6a6a67 |
| institution | Directory Open Access Journal |
| issn | 2191-0855 |
| language | English |
| last_indexed | 2024-04-12T03:53:25Z |
| publishDate | 2017-06-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj.art-f0a6ffdaff8f44529334e4d42c6a6a672022-12-22T03:48:55ZengSpringerOpenAMB Express2191-08552017-06-017111210.1186/s13568-017-0400-7A novel constructed SPT15 mutagenesis library of Saccharomyces cerevisiae by using gTME technique for enhanced ethanol productionAshraf A. M. M. El-Rotail0Liang Zhang1Youran Li2Shuang Ping Liu3Gui Yang Shi4The Key Laboratory of Industrial Biotechnology, Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan UniversityThe Key Laboratory of Industrial Biotechnology, Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan UniversityThe Key Laboratory of Industrial Biotechnology, Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan UniversityThe Key Laboratory of Industrial Biotechnology, Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan UniversityThe Key Laboratory of Industrial Biotechnology, Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan UniversityAbstract During the last few years, the global transcription machinery engineering (gTME) technique has gained more attention as an effective approach for the construction of novel mutants. Genetic strategies (molecular biology methods) were utilized to get mutational for both genes (SPT15 and TAF23) basically existed in the Saccharomyces cerevisiae genome via screening the gTME approach in order to obtain a new mutant S. cerevisiae diploid strain. The vector pYX212 was utilized to transform these genes into the control diploid strain S. cerevisiae through the process of mating between haploids control strains S. cerevisiae (MAT-a [CICC 1374]) and (MAT-α [CICC 31144]), by using the oligonucleotide primers SPT15-EcoRI-FW/SPT15-SalI-RV and TAF23-SalI-FW/TAF23-NheI-RV, respectively. The resultant mutants were examined for a series of stability tests. This study showed how strong the effect of using strategic gTME with the importance of the modification we conducted in Error Prone PCR protocol by increasing MnCl2 concentration instead of MgCl2. More than ninety mutants we obtained in this study were distinguished by a high level production of bio-ethanol as compared to the diploid control strain.http://link.springer.com/article/10.1186/s13568-017-0400-7BioethanolError-prone PCREthanol productionEthanol toleranceGlobal transcription machinery engineeringSPT15 |
| spellingShingle | Ashraf A. M. M. El-Rotail Liang Zhang Youran Li Shuang Ping Liu Gui Yang Shi A novel constructed SPT15 mutagenesis library of Saccharomyces cerevisiae by using gTME technique for enhanced ethanol production AMB Express Bioethanol Error-prone PCR Ethanol production Ethanol tolerance Global transcription machinery engineering SPT15 |
| title | A novel constructed SPT15 mutagenesis library of Saccharomyces cerevisiae by using gTME technique for enhanced ethanol production |
| title_full | A novel constructed SPT15 mutagenesis library of Saccharomyces cerevisiae by using gTME technique for enhanced ethanol production |
| title_fullStr | A novel constructed SPT15 mutagenesis library of Saccharomyces cerevisiae by using gTME technique for enhanced ethanol production |
| title_full_unstemmed | A novel constructed SPT15 mutagenesis library of Saccharomyces cerevisiae by using gTME technique for enhanced ethanol production |
| title_short | A novel constructed SPT15 mutagenesis library of Saccharomyces cerevisiae by using gTME technique for enhanced ethanol production |
| title_sort | novel constructed spt15 mutagenesis library of saccharomyces cerevisiae by using gtme technique for enhanced ethanol production |
| topic | Bioethanol Error-prone PCR Ethanol production Ethanol tolerance Global transcription machinery engineering SPT15 |
| url | http://link.springer.com/article/10.1186/s13568-017-0400-7 |
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