Abstract
This chapter discusses the processes of immunological detection and characterization of products translated from cloned deoxyribonucleic acid (DNA) fragments. The most general assay for screening bacterial clones for gene products encoded by cloned foreign DNA is based on the antibody-mediated recognition of antigenic determinants. Such an immunoassay is capable of detecting incompletely translated products as well as proteins that have no easily detectable or selectable functions. The chapter describes a simple and sensitive solid-phase indirect radioimmunoassay developed for the detection of an antigen in liquid cultures or in situ in bacterial colonies and phage plaques; this radioimmunoassay is capable of identifying an antigen in polyacrylamide gels, thereby making possible the electrophoretic characterization of translation products of foreign gene fragments cloned into plasmid or phage vectors. The assay that can be carried out using either derivatized cellulose filters or polyvinyl sheets as a solid-phase matrix is used to identify and characterize an electrophoretically mouse dihydrofolate reductase (DHFR) antigen in Escherichia coli (E. coli) cells containing a chimeric plasmid coding for the mouse DHFR.
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