Cloning and Expression of Bombyx mori Silk Gland Elongation Factor 1γ in Escherichia coli

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2002, Vol.66 (3), p.558-565
• Main Authors: KAMIIE, Katsuyoshi, NOMURA, Yoshitaka, KOBAYASHI, Satoru, TAIRA, Hideharu, KOBAYASHI, Kohmei, YAMASHITA, Tetsuro, KIDOU, Shin-ichiro, EJIRI, Shin-ichiro
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Bombyx mori; elongation factor 1γ; Escherichia coli; Fundamental and applied biological sciences. Psychology; glutathione; glutathione S-transferase (GST); Molecular and cellular biology; Molecular genetics; Translation. Translation factors. Protein processing
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.66.558

Elongation factor 1 (EF-1) from the silk gland of Bombyx mori consists of α-, β-, γ-, and δ-subunits. EF-1α*GTP catalyzes the binding of aminoacyl-tRNA to ribosomes concomitant with the hydrolysis of GTP. EF-1βγδ catalyzes the exchange of EF-1α-bound GDP for exogenous GTP and stimulates the EF-1α-dependent binding of aminoacyl-tRNA to ribosomes. EF-1γ 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-1γ shares 67.3% amino acid identity with Artemia salina EF-1γ. The N-terminal domain (amino acid residues 1-211) of silk gland EF-1γ is 29.3% identical to maize glutathione S-transferase. We demonstrated that silk gland EF-1γ bound to glutathione Sepharose, suggesting that the N-terminal domain of EF-1γ may have the capacity to bind to glutathione.