Cloning and Expression ofBombyx moriSilk Gland Elongation Factor 1γ inEscherichia coli
| Autor: | Shin-ichiro Ejiri, Shin-ichiro Kidou, Yoshitaka Nomura, Kohmei Kobayashi, Hideharu Taira, Katsuyoshi Kamiie, Tetsuro Yamashita, Satoru Kobayashi |
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| Rok vydání: | 2002 |
| Předmět: |
DNA
Complementary GTP' Molecular Sequence Data Applied Microbiology and Biotechnology Biochemistry Analytical Chemistry Bombycidae Exocrine Glands Peptide Elongation Factor 1 Bombyx mori Complementary DNA Escherichia coli Animals Amino Acid Sequence Cloning Molecular Molecular Biology Conserved Sequence Glutathione Transferase chemistry.chemical_classification Base Sequence biology Reverse Transcriptase Polymerase Chain Reaction cDNA library fungi Organic Chemistry General Medicine Bombyx Chromatography Ion Exchange biology.organism_classification Glutathione Molecular biology Amino acid Elongation factor Open reading frame chemistry Electrophoresis Polyacrylamide Gel Biotechnology |
| Zdroj: | Bioscience, Biotechnology, and Biochemistry. 66:558-565 |
| ISSN: | 1347-6947 0916-8451 |
| DOI: | 10.1271/bbb.66.558 |
| Popis: | Elongation factor 1 (EF-1) from the silk gland of Bombyx mori consists of alpha-, beta-, gamma-, and delta-subunits. EF-1alpha GTP catalyzes the binding of aminoacyl-tRNA to ribosomes concomitant with the hydrolysis of GTP. EF-1betagammadelta catalyzes the exchange of EF-1alpha-bound GDP for exogenous GTP and stimulates the EF-1alpha-dependent binding of aminoacyl-tRNA to ribosomes. EF-1gamma cDNA, which contains an open reading frame (ORF) encoding a polypeptide of 423 amino acid residues, was amplified and cloned by PCR from a silk gland cDNA library. The calculated molecular mass and predicted pI of the product were 48,388 Da and 5.84, respectively. The silk gland EF-1gamma shares 67.3% amino acid identity with Artemia salina EF-lgamma. The N-terminal domain (amino acid residues 1-211) of silk gland EF-lgamma is 29.3% identical to maize glutathione S-transferase. We demonstrated that silk gland EF-lgamma bound to glutathione Sepharose, suggesting that the N-terminal domain of EF-1gamma may have the capacity to bind to glutathione. |
| Databáze: | OpenAIRE |
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