Cloning and sequencing of a cDNA encoding glyceraldehyde-3-phosphate dehydrogenase from Tetrahymena thermophila: growth-associated changes in its mRNA expression
| Autor: | Yutong Zhao, Yoshinori Nozawa, Shigeru Nakashima, Masataka Andoh |
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| Rok vydání: | 1997 |
| Předmět: |
DNA
Complementary Genes Protozoan Molecular Sequence Data Molecular cloning Microbiology Gene Expression Regulation Enzymologic Tetrahymena thermophila Complementary DNA Gene expression Animals Northern blot Amino Acid Sequence RNA Messenger Cloning Molecular Glyceraldehyde 3-phosphate dehydrogenase biology cDNA library Tetrahymena Sequence Analysis DNA DNA Protozoan biology.organism_classification Molecular biology Open reading frame Biochemistry biology.protein RNA Protozoan |
| Zdroj: | The Journal of eukaryotic microbiology. 44(5) |
| ISSN: | 1066-5234 |
| Popis: | Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in the glycolytic pathway. Since its transcript levels do not vary in most experimental conditions, it has been often used as a control in northern blot or reverse transcriptase-polymerase chain reaction analysis. We have cloned and sequenced a gene encoding glyceraldehyde-3-phosphate dehydrogenase (Tthgapdh) from Tetrahymena thermophila cDNA library and determined whether the Tthgapdh mRNA is a loading control in gene expression studies of T. thermophila cell. The open reading frame encoded a protein of 341 amino acid residues (36.8 kDa) containing a nicotinamide adenine dinucleotide-binding domain and a catalytic domain, which was highly similar to those of other organisms. Its mRNA levels at different growth stages were examined by northern blot analysis. The fragment of the isolated cDNA was hybridized to a 1.3-kb mRNA transcript. There was a marked increase in Tthgapdh mRNA level at the mid-exponential phase, followed by a gradual decrease. Therefore, much caution should be made to use Tthgapdh mRNA as an internal standard for northern blot analysis in Tetrahymena. |
| Databáze: | OpenAIRE |
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