Functional cloning

<p>Functional cloning is a <a href="/facts/Molecular_cloning/Te56373e">molecular cloning</a> technique that relies on prior knowledge of the encoded <a href="/facts/Protein/Jxmagu3E">protein</a>’s <a href="/facts/Protein_sequence/QNBoERdu">sequence</a> or <a href="/facts/Protein_function/Jxmagu3E">function</a> for <a href="/facts/Gene/e1swwPY6">gene</a> identification. In this assay, a <a href="/facts/Genomic_library/NnE0fPVC">genomic</a> or <a href="/facts/CDNA_library/MPMUmT0w">cDNA library</a> is screened to identify the genetic sequence of a protein of interest. <a href="/facts/Expression_vector/yqjSkTgE">Expression</a> cDNA libraries may be screened with <a href="/facts/Antibody/MX8pdTdP">antibodies</a> 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 <a href="/facts/Hybridization_probe/j3pu1L7E">probed</a> against the library to identify the gene encoding the protein of interest. Once candidate clones carrying the gene of interest are identified, they are <a href="/facts/DNA_sequencing/hsPsNCQW">sequenced</a> and their identity is confirmed. This method of cloning allows researchers to screen entire <a href="/facts/Genome/31Sm08JT">genomes</a> without prior knowledge of the location of the gene or the genetic sequence.
</p><p>This technique can be used to identify genes that encode similar proteins from one organism to another. Similarly, this technique can be paired with <a href="/facts/Metagenomics/GWFeMd7z">metagenomic</a> libraries to identify novel genes and proteins that perform similar functions, such as the identification of novel <a href="/facts/Antibiotics/sF1kIfxg">antibiotics</a> by screening for <a href="/facts/Beta-lactamase/8PdHT21m">beta-lactamase</a> activity or selecting for growth in the presence of <a href="/facts/Penicillin/Abo8VQ3p">penicillin</a>.
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Functional cloning open-in-new

Functional cloning