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Cloning and characterization of the cDNA coding for the catalytic α subunit of CK2 from tobacco

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Part of the book series: Molecular and Cellular Biochemistry ((DMCB,volume 35))

Abstract

We have previously reported the participation of the protein kinase CK2 in the mechanism by which salicylic acid activates transcription of genes, such as those coding for glutathion S-transferases, in tobacco. With the purpose of further studying the participation of CK2 in this signal transduction pathway, we isolated and sequenced the cDNA from the NtCK2A gene, coding for the catalytic a subunit of CK2 from tobacco. The NtCK2A cDNA was isolated by screening of a tobacco cDNA library with a heterologous probe from Arabidopsis thaliana, followed by 3’ RACE to obtain the 3’ region. Sequence analysis of the NtCK2A cDNA showed a high level of identity between this CK2a protein sequence and the corresponding sequences of other plant species such as Arabidopsis and maize (92-95% identity), or those of animal species such as human and Xenopus laevis (75% identity). The expression of the NtCK2A gene in different tissues from tobacco plants was analyzed by Northern blot. High levels of expression of this gene were observed in proliferating tissues such as shoot and root apical meristems. A recombinant CK2α protein was obtained after expression of the NtCK2A cDNA in Escherichia coli. The ability of this recombinant CK2α subunit to phosphorylate casein was inhibited by heparin and stimulated by the CK2β subunit from Xenopus laevis. (Mol Cell Biochem 227: 129-135, 2001)

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Salinas, P., Bantignies, B., Tapia, J., Jordana, X., Holuigue, L. (2001). Cloning and characterization of the cDNA coding for the catalytic α subunit of CK2 from tobacco. In: Ahmed, K., Issinger, OG., Allende, J.E. (eds) Protein Kinase CK2 — From Structure to Regulation. Molecular and Cellular Biochemistry, vol 35. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-1723-8_16

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  • DOI: https://doi.org/10.1007/978-1-4615-1723-8_16

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-5696-7

  • Online ISBN: 978-1-4615-1723-8

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