Workflow for utilizing selection in a functional cloning experiment. Here only genes providing ampicillin resistance are selected.
Functional cloning is a molecular cloning technique that relies on prior knowledge of the encoded protein’s sequence or function for gene identification.[1][2][3] In this assay, a genomic or cDNA library is screened to identify the genetic sequence of a protein of interest. Expression cDNA libraries may be screened with antibodies specific for the protein of interest or may rely on selection via the protein function.[1] Historically, the amino acid sequence of a protein was used to prepare degenerate oligonucleotides which were then probed against the library to identify the gene encoding the protein of interest.[2][3] Once candidate clones carrying the gene of interest are identified, they are sequenced and their identity is confirmed. This method of cloning allows researchers to screen entire genomes without prior knowledge of the location of the gene or the genetic sequence.[1]
This technique can be used to identify genes that encode similar proteins from one organism to another.[4] Similarly, this technique can be paired with metagenomic libraries to identify novel genes and proteins that perform similar functions, such as the identification of novel antibiotics by screening for beta-lactamase activity or selecting for growth in the presence of penicillin.[5]
^ abcLam, Kathy N.; Cheng, Jiujun; Engel, Katja; Neufeld, Josh D.; Charles, Trevor C. (2015). "Current and future resources for functional metagenomics". Frontiers in Microbiology. 6: 1196. doi:10.3389/fmicb.2015.01196. ISSN 1664-302X. PMC 4625089. PMID 26579102.
^ abOxford dictionary of biochemistry and molecular biology. Cammack, Richard, Ph. D. (Rev. ed.). Oxford: Oxford University Press. 2006. ISBN 9780198529170. OCLC 65467611.{{cite book}}: CS1 maint: others (link)
^ abSchindler-Hoehn; Hoehn, Holger (2007-01-01). Fanconi Anemia: A Paradigmatic Disease for the Understanding of Cancer and Aging. Karger Medical and Scientific Publishers. ISBN 9783805582773.
^Skjøt, M.; Kauppinen, S.; Kofod, L.; Fuglsang, C.; Pauly, M.; Dalbøge, H.; Andersen, L. (2001-07-01). "Functional cloning of an endo-arabinanase from Aspergillus aculeatus and its heterologous expression in A. oryzae and tobacco". Molecular Genetics and Genomics. 265 (5): 913–921. doi:10.1007/s004380100489. ISSN 1617-4615. PMID 11523809. S2CID 20314288.
^Allen, Heather K; Moe, Luke A; Rodbumrer, Jitsupang; Gaarder, Andra; Handelsman, Jo (2008-10-09). "Functional metagenomics reveals diverse β-lactamases in a remote Alaskan soil". The ISME Journal. 3 (2): 243–251. doi:10.1038/ismej.2008.86. ISSN 1751-7370. PMID 18843302.
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(BAC) is a DNA construct, based on a functional fertility plasmid (or F-plasmid), used for transforming and cloning in bacteria, usually E. coli. F-plasmids...
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clone manufacturers have been challenged in court by Lego. The lawsuits have been mostly unsuccessful, for courts have generally found the functional...
Bennett MK, Molineaux S, Luo Y, Payan DG, Mancebo HS, Wu J (Nov 2001). "Functionalcloning of Src-like adapter protein-2 (SLAP-2), a novel inhibitor of antigen...
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Virkkala. It was born as a functionalclone of RemoteAccess BBS (which in turn was a clone of QuickBBS), but extended the functionality with several newer technology...
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