A novel constructed SPT15 mutagenesis library of Saccharomyces cerevisiae by using gTME technique for enhanced ethanol production
| Autor: | Gui Yang Shi, Liang Zhang, Ashraf A. M. M. El-Rotail, Shuang Ping Liu, Youran Li |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine lcsh:Biotechnology Saccharomyces cerevisiae Mutant Biophysics lcsh:QR1-502 Bioethanol Ethanol tolerance Biology MOLECULAR BIOLOGY METHODS 01 natural sciences Applied Microbiology and Biotechnology Genome lcsh:Microbiology 03 medical and health sciences Transcription (biology) 010608 biotechnology lcsh:TP248.13-248.65 Ethanol fuel Global transcription machinery engineering Gene Genetics Error-prone PCR biology.organism_classification 030104 developmental biology Ethanol production SPT15 Original Article Ploidy |
| Zdroj: | AMB Express, Vol 7, Iss 1, Pp 1-12 (2017) AMB Express |
| ISSN: | 2191-0855 |
| DOI: | 10.1186/s13568-017-0400-7 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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