Cloning and characterization of two alleles of the murine extracellular superoxide dismutase gene

Andrej Mirossay, Sujung Jun, Ladislav Dory

Research output: Contribution to journalArticle

Abstract

We have recently documented the existence of a second allele of ecSOD in mice. Thus far, this allele was only found in the 129P3/J strain. It is characterized by two point mutations leading to amino acid changes as well as a 10 bp deletion from the 3′ UTR. We have also shown that the phenotype is profoundly affected by the genotype. In order to obtain a tool to investigate the differences in the properties as well as the posttranscriptional regulation of expression of the two alleles we now describe the creation and characterization of stably transfected CHO-K1 cell lines expressing either of these alleles. CHO-K1 cells were chosen because they do not express endogenous ecSOD and are easy to transfect. We demonstrate that the transfected cells secrete substantial amounts of glycosylated ecSOD, detected by Western blot analyses, ConA-Sepharose affinity chromatography and activity measurements.

Original languageEnglish (US)
Pages (from-to)739-743
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume352
Issue number3
DOIs
StatePublished - Jan 19 2007
Externally publishedYes

Fingerprint

Cloning
Superoxide Dismutase
Organism Cloning
Genes
Alleles
Affinity chromatography
CHO Cells
3' Untranslated Regions
Sepharose
Amino Acids
Agarose Chromatography
Affinity Chromatography
Point Mutation
Western Blotting
Genotype
Phenotype
Cell Line

Keywords

  • CHO cells
  • Cloning
  • ecSOD expression
  • Extracellular superoxide dismutase
  • Mouse

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Cloning and characterization of two alleles of the murine extracellular superoxide dismutase gene. / Mirossay, Andrej; Jun, Sujung; Dory, Ladislav.

In: Biochemical and Biophysical Research Communications, Vol. 352, No. 3, 19.01.2007, p. 739-743.

Research output: Contribution to journalArticle

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