RRC ID 73956
Author Inouye S.
Title Cloning, expression, purification and characterization of an isotype of clytin, a calcium-binding photoprotein from the luminous hydromedusa Clytia gregarium.
Journal J Biochem
Abstract The cDNA for an isotype of clytin, a calcium-binding photoprotein from the luminous jellyfish Clytia gregarium, was identified and named clytin-II. The histidine-tagged apoprotein of clytin-II expressed into the periplasmic space of Escherichia coli cells was isolated by nickel chelate affinity chromatography. Recombinant clytin-II regenerated from apoprotein by incubation with coelenterazine was purified. The yield of purified clytin-II was 13 mg from 2 l of cultured cells with purity >95%. The luminescence properties of clytin-II were characterized by comparison with clytin-I and aequorin, and semi-synthetic clytin-II was also prepared. The initial luminescence intensity of clytin-II triggered by Ca(2+) was 4.5 times higher than that of clytin-I and aequorin, but the luminescence capacity was close to clytin-I and aequorin. Thus, clytin-II is a useful protein, showing high sensitivity in the signal-to-noise ratio of luminescence intensity.
Volume 143(5)
Pages 711-7
Published 2008-5-1
DOI 10.1093/jb/mvn024
PII mvn024
PMID 18296715
MeSH Aequorin / immunology Amino Acid Sequence Animals Antibodies Apoproteins / immunology Calcium / metabolism Calcium-Binding Proteins / chemistry* Calcium-Binding Proteins / genetics* Calcium-Binding Proteins / metabolism Cloning, Molecular Escherichia coli / genetics Hydrozoa / chemistry Hydrozoa / genetics* Immunoblotting Luminescent Proteins / chemistry* Luminescent Proteins / genetics* Luminescent Proteins / metabolism Molecular Sequence Data Recombinant Proteins / chemistry Recombinant Proteins / immunology Recombinant Proteins / isolation & purification Recombinant Proteins / metabolism Sequence Homology, Amino Acid
IF 2.476
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The most frequently cited source Patent(IFI CLAIMS)
Total number of mentions 12
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Resource
DNA material pCL31 (RDB19838)