Document Details
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Document Type |
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Article In Journal |
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Document Title |
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CHARACTERIZATION OF GLYCEROL-3- PHOSPHATE DEHYDROGENASE (GPD1) GENE FROM EGYPTIAN YEAST WITH FLUORESCENCE DIFFERENTIAL DISPLAY CHARACTERIZATION OF GLYCEROL-3- PHOSPHATE DEHYDROGENASE (GPD1) GENE FROM EGYPTIAN YEAST WITH FLUORESCENCE DIFFERENTIAL DISPLAY |
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Subject |
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Biological |
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Document Language |
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English |
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Abstract |
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The main task of the present work was to isolate and characterize abiotic stress-induced gene(s) from yeast (Sac-charomyces cerevisiae) through fluores-cence differential display-polymerase chain reaction (fluorescence DD-PCR). Six Egyptian isolates of yeast were evaluated under salt and osmotic stresses at concentrations of 2 M NaCl and 2 M sorbitol, respectively, in which one isolate was pre selected as the most tolerant against both stresses. Fluorescence DD-PCR was conducted for cDNAs after being exposed to 0 and 2 M NaCl for 0, 20, 40 and 60 minutes. After a number of DD-PCR runs, 350 fragments were observed, 30 of them (9.14%) showed differential expression versus exposure times. They were classified into 12 patterns of gene expression. Three up-regulated DD fragments, above 100 bp in size, were chosen and cloned for subsequent molecular analysis and gene construction. DNA sequences were detected and then subjected to homology searching via computer software. The results indicated that one DD fragment showed significant homology with a yeast DNA fragment glycerol-3-phosphate dehydrogenase (GPD1); a NAD dependent key enzyme of glycerol synthesis that is essential for growth under osmotic stress. This gene fragment was chosen as the candidate gene to recover full-length DNA following the rapid amplification of cDNA ends (RACE) strategy. The full-length GPD1 gene was cloned and transformed into E. coli. The results of stress tolerance for GPD1-transformed bacteria indicated the efficacy of utilizing the gene in conferring salt tolerance at the prokaryotic level. |
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ISSN |
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0046-161X |
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Journal Name |
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Egyptian Journal of Genetics and Cytology |
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Volume |
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38 |
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Issue Number |
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1 |
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Publishing Year |
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2009 AH
2009 AD |
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Article Type |
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Article |
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Added Date |
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Monday, March 22, 2010 |
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Researchers
| شريف إدريس | EDRIS, Sheriff | Researcher | Doctorate | |
| أحمد بهي الدين | BAHIELDIN, Ahmad | Researcher | Doctorate | |
| فتوح محمد الدمياطي | DOMYATI, Falleh M | Researcher | Doctorate | fm_domyati@hotmail.com |
| رانية أحمد يونس | YOUNIS, RANIA Ahmad | Researcher | Doctorate | |
| سيد حسن حسنين | HASSANEIN, S H | Researcher | Doctorate | |
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