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Journal Article

Detection of Proteins like Human $\gamma$ and $\beta$ Globins in Escherichia coli Carrying Recombinant DNA Plasmids

Lois B. Wilson, Hernan E. Grenett, Daniel M. Doran, Fred A. Garver and John T. Wilson
Proceedings of the National Academy of Sciences of the United States of America
Vol. 76, No. 11 (Nov., 1979), pp. 5631-5635
Stable URL: http://www.jstor.org/stable/70511
Page Count: 5
Escherichia coli strain $\chi$1776 carrying recombinant DNA plasmids containing cDNA copies of human $\beta$ or $\gamma$ globin mRNAs has been shown by radioimmunoassay to synthesize polypeptides antigenically related to the $\beta$ and $\gamma$ chains of human hemoglobin. The $\gamma$ and $\beta$ polypeptides have been enriched from lysates on immunoabsorbent columns containing hemoglobin antibodies and shown to specifically inhibit the antigen-antibody binding between $^{125}$I-labeled hemoglobin and the homologous antibody but not other hemoglobin-antihemoglobin reactions. Clone JW151, which is known to contain a complete copy of the coding portion of the $\gamma$ globin mRNA, has been shown to produce a protein that reacts specifically with antibody to the $\gamma$ chain of fetal hemoglobin, hemoglobin Kenya, and hemoglobin Bart's.