Screening λgt11 Expression Libraries with Antibody Probes
Antibodies can be used as probes to isolate directly a particular gene of interest. A recombinant DNA library is first constructed in an Escherichia coli expression vector, and foreign antigens are expressed in E. coli from the DNA inserts. The antigens produced are transferred onto nitrocellulose filters and then probed with antibody to detect the desired recombinant. This approach is rapid, and is particularly useful for isolating genes that are expressed at a low level and therefore are difficult to isolate using techniques employing nucleic acid probes (see Ref. 1) or by hybrid/selection methods.2 This chapter describes methods for isolating eukaryotic genes by constructing and screening E. coli expression vector libraries with antibody probes using the bacteriophage expression vector λgt11.3,4
KeywordslacZ Gene Expression Library Antibody Probe locI Plasmid Overlay Plate
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