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A murine Thy-1.2 reporter vector containing a SV40 origin for rapid cloning and analysis of eukaryotic promoters
Y Kadokawa1, T Kusakabe, Y Kamachi
1Division of Molecular Biology, Meiji Institute of Health Science, Odawara, Japan.
Abstract:
A new vector, pATO, was constructed for rapid cloning and analysis of eukaryotic promoters. When a recombinant pATO, carrying a promoter sequence in its multiple cloning site, was introduced into COS cells, Thy-1.2 protein was produced on the cell surface, and was easily identified by an fluorescein-conjugated anti-Thy-1.2 antibody. The intensity of the fluorescence reflected the strength of the inserted promoter. Since pATO could replicate efficiently in COS cells, the recombinant plasmids recovered from a single COS cell were sufficient to transform Escherichia coli cells. This plasmid is applicable for the rapid and labor saving cloning of promoter elements.