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Phage display cycle. 1) fusion proteins for a viral coat protein + the gene to be evolved (typically an antibody fragment) are expressed in bacteriophage. 2) the library of phage are washed over an immobilised target. 3) the remaining high-affinity binders are used to infect bacteria. 4) the genes encoding the high-affinity binders are isolated.
New England Biolabs (NEB) is an American life sciences company which produces and supplies recombinant and native enzyme reagents for life science research. [2] It also provides products and services supporting genome editing , synthetic biology and next-generation sequencing . [ 3 ]
Plaque exhibiting bacterial lawn with clearings made by Artharobacter phage GantcherGoblin.. The Actinobacteriophage database, more commonly known as PhagesDB, is an interactive, comprehensive, database-backed website that collects and shares information related to the discovery, characterization and genomics of viruses that typically infect Actinobacterial hosts.
Phage display is a different use of phages involving a library of phages with a variable peptide linked to a surface protein. Each phage genome encodes the variant of the protein displayed on its surface (hence the name), providing a link between the peptide variant and its encoding gene.
The end result is the peptides produced by bacteriophage are specific. The resulting filamentous phages can infect gram-negative bacteria once again to produce phage libraries. The cycle can occur many times resulting with strong affinity binding peptides to the target.
A genomic library is a collection of overlapping DNA fragments that together make up the total genomic DNA of a single organism. The DNA is stored in a population of identical vectors , each containing a different insert of DNA.
Virulent Phage (Selected Papers in Biochemistry, Volume 1). University of Tokyo Press, Tokyo, and University Park Press, Baltimore, MD. OCLC 208390, ISBN 0-8391-0612-2
In biotechnology applications, T7 RNA polymerase is commonly used to transcribe DNA that has been cloned into vectors that have two (different) phage promoters (e.g., T7 and T3, or T7 and SP6) in opposite orientation. RNA can be selectively synthesized from either strand of the insert DNA with the different polymerases.