Abstract
Conventional procedures to isolate a gene belonging to an ortholog family usually imply the use or the construction of double-stranded cDNA libraries derived from a specific mRNA source of interest (cells or tissues) (1). The double-stranded DNA library plated on various membranes is screened by filter hybridization with a radiolabeled probe (1) derived from the known homologous gene. Clones or plaques hybridizing with the probe are then isolated and sequenced to find the gene of interest. This approach is time-consuming and a large number of false positive clones might be obtained, given that no homology is a priori available, especially when the homologous probe contains a short and stable stretch of a sequence sharing clustered homology with both strands of the cDNA library. Alternatively, homologous genes may be isolated by PCR, but only when specific primers are available.
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Mastrangeli, R., Donini, S. (2002). Cloning of Homologous Genes by Gene-Capture PCR. In: Chen, BY., Janes, H.W. (eds) PCR Cloning Protocols. Methods in Molecular Biology™, vol 192. Humana Press. https://doi.org/10.1385/1-59259-177-9:359
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DOI: https://doi.org/10.1385/1-59259-177-9:359
Publisher Name: Humana Press
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Online ISBN: 978-1-59259-177-0
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