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Functional Cloning Using,,,ViraPort Retroviral cDNA Expression Libraries

Phenotypic complementation

Signaling proteins

Loss of contact-inhibition

Phenotypic complementation

Reporter activation

Transcription factors

Reporter activation

Apoptosis inhibitors

Survival: resistance to apoptosis inducers

Metastasis-inducing genes

In vivo metastasis

In vitro invasion

Differentiation-inducing genes

Phenotype: differentiation

Ion channels

Phenotype: ion-specific indicator or tracer

Expression cloning allows genes to be identified and isolated based on gene function or phenotype in the absence of any prior knowledge of protein or nucleic acid sequence. A wide range of gene types have been cloned from complex cDNA libraries using functional assays that allow a specific trait to be selected or screened (Table 1).1 Originally, hybrid selection, direct transfection of plasmid expression libraries, and transfection of genomic DNA were successfully used to clone growth factors, cell surface proteins, and transcription factors. However, selected cells or cDNA pools usually contained a large number (often hundreds) of extraneous clones in addition to the target cDNA of interest, necessitating several rounds of laborious DNA isolation
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