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Investigation of neuronal cell type-specific gene expression of Ca2+/calmodulin-dependent protein kinase II

Abstract

The promoter activity of the rat Ca2+/calmodulin-dependent protein kinase II gene was analyzed using the luciferase reporter gene in neuronal and non-neuronal cell lines. Neuronal cell type-specific promoter activity was found in the 5′-flanking region of α and β isoform genes of the kinase. Silencer elements were also found further upstream of promoter regions. A brain-specific protein bound to the DNA sequence of the 5′-flanking region of the gene was found by gel mobility shift analysis in the nuclear extract of the rat brain, including the cerebellum, forebrain, and brainstem, but not in that of non-neuronal tissues, including liver, kidney and spleen. The luciferase expression system and gel shift analysis can be used as an additional and better index by which to monitor gene expression in most cell types.

Abbreviations

CaM kinase II:

Ca2+/calmodulin-dependent protein kinase II

CPRG:

chlorophenol red β-D-galactopyranoside

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

fetal bovine serum

EGTA:

ethylene glycol bis (β-aminoethylether)- N,N,N′,N′-tetra acetic acid

PAGE:

polyacrylamide gel electrophoresis

PBS:

phosphate buffered saline

PCR:

polymerase chain reaction

PMSF:

phenylmethyl sulfonyl fluoride

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Correspondence to Takashi Yamauchi.

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Published: April 12, 2002

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Mima, K., Donai, H. & Yamauchi, T. Investigation of neuronal cell type-specific gene expression of Ca2+/calmodulin-dependent protein kinase II. Biol Proced Online 3, 79–89 (2001). https://doi.org/10.1251/bpo26

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Indexing terms

  • Ca2+/calmodulin-dependent protein kinase II (CaM kinase II, CaMKII)
  • gene expression
  • promoter activity
  • silencer element
  • neuronal cell
  • gel shift analysis