2004
DOI: 10.1080/10425170310001652174
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Molecular Cloning of Bovine eIF5A and Deoxyhypusine Synthase cDNA

Abstract: Deoxyhypusine synthase is the first of the two enzymes that catalyzes the maturation of eukaryotic initiation factor 5A (eIF5A). The mature eIF5A is the only known protein in eukaryotic cells that contains the unusual amino acid hypusine (N(epsilon)-(4-amino-2(R)-hydroxybutyl)-lysine). Synthesis of hypusine is essential for the function of eIF5A in eukaryotic cell proliferation and survival. Here we describe the cloning and characterization of bovine eIF5A and bovine deoxyhypusine synthase. The deduced bovine … Show more

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Cited by 5 publications
(2 citation statements)
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“…The desired bands were gel purified, cloned into pET15b (digested with XhoI and dephosphorylated), and transformed into JM109 competent cells. The transformants were selected by ampicillin resistance followed by Quick Screening [68]. The orientation of the insert in the selected colonies was checked by colony PCR as described before.…”
Section: Pcr Amplification and Cloning Of Putative 2 O -Adh Genementioning
confidence: 99%
“…The desired bands were gel purified, cloned into pET15b (digested with XhoI and dephosphorylated), and transformed into JM109 competent cells. The transformants were selected by ampicillin resistance followed by Quick Screening [68]. The orientation of the insert in the selected colonies was checked by colony PCR as described before.…”
Section: Pcr Amplification and Cloning Of Putative 2 O -Adh Genementioning
confidence: 99%
“…Each ligation reaction was then transformed into JM109 competent cells according to the method of Chung et al (Chung et al, 1989) and plated on ampicillin containing plates. The transformants were screened to find recombinant DNA containing colonies by Quick Screening (Huang et al, 2004). In Quick Screening, a toothpick was used to isolate a colony and the cells were resuspended in 25 μl of STE solution (100 mM NaCl, 20 mM Tris-Cl pH 7.5, and 10 mM EDTA).…”
Section: Primersmentioning
confidence: 99%