page_title: Functional cloning
hierarchical_section_title: Functional cloning
caption_attribution_description: English: Workflow for utilizing selection in a functional cloning experiment. Here only genes providing ampicillin resistance are selected.
context_page_description: Functional cloning is a molecular cloning technique that relies on prior knowledge of the encoded protein’s sequence or function for gene identification. 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. 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. 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.
  This technique can be used to identify genes that encode similar proteins from one organism to another.
context_section_description: Functional cloning is a molecular cloning technique that relies on prior knowledge of the encoded protein’s sequence or function for gene identification. 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. 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. 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.
  This technique can be used to identify genes that encode similar proteins from one organism to another. 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.
caption: Workflow for utilizing selection in a functional cloning experiment. Here only genes providing ampicillin resistance are selected.
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