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The video is uploaded only to provide the download linkA human monoclonal antibody to human gamma-globin gene mRNA directs the synthesis of a precursor of gamma-globin protein in a human cell line.
We have produced a human monoclonal antibody (MAb) to gamma-globin mRNA by using the technique of immunoglobulin-gene transfection. The MAb 1D6, which has a heavy chain of IgG1 and light chain of kappa, was used to affinity-purify membrane fractions from a human erythroleukemia cell line (K562) and fluorescein isothiocyanate-labeled mRNA for affinity purification. The membrane fraction bound mRNA by a process thought to be specific because the MAb could not cross react with other mRNAs tested. In immuno-precipitation experiments, the membrane fraction bound to the protein products of the isolated gamma-globin mRNA. The MAb 1D6 also inhibited the synthesis of the protein product in a cell-free translation system using human gamma-globin mRNA in addition to the recombinant ribosome. The inhibition of the synthesis of the protein product in the cell-free system required the intact translation initiation signal of the mRNA. These data support the notion that the MAb 1D6 recognizes the translation initiation signal of human gamma-globin mRNA. In addition, the gene product of the mRNA might be synthesized as a precursor of a gamma-globin protein. These results are the first demonstration of the gene-specific translation of a naturally occurring mRNA in a eukaryotic translation system.
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