User:Pakpoom Subsoontorn/Notebook/Genetically Encoded Memory/2008/10/28

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PhiC31
length:: 613 aa crystal structure is unknown.
 * Host: Streptomyces lividans
 * Action: tyrosine integrase
 * Structure:
 * Target sequence: attB:: has two inverted repeated sequence (core-type) flanking an overlap region (TTG). Minimal attB is 34 bps while minimal attP is 39 bp for e.coli intramolecular integration. attP:: has core-type sites and overlap region, without an arm-type site. --See the schematic sequence of attB and attP at (1-f1). See Groth-2000 for experiment for minimal sequence. --See Thyagarajan-2001 for integration of attB and attP above background/pseudointegration site ~attP in human genome (these could be very cool model experiment!)

Mutation/directed evolution could change specificity to pseudo attP site (HpsA, 44% identity) Excisionase has not yet been found.
 * Change specificity:
 * Mechanism:
 * Excision:


 * mutant/hybrid
 * directed evolution for targeting human's site


 * Non-natural host:
 * Integration works in E.coli, mammalian and in vitro,
 * Mouse embryos Hollis et al 2003
 * Bovine fiboblase--Ma et al 2006
 * Drosophila--Groth et al 2004
 * Aedes aegyptus--Nimmo et al 2006
 * Silk worm--Nakayama et al 2006
 * cassette exchange: Drosphila --Bateman et al 2006, fission yeast--Thomson et al 2001, mouse embryonic stem cell--Belteki et al 2003
 * Xenopus laevis--Allen&Weeks 2005
 * chloroplast transformation--Lutz et al 2004
 * gene therapy example--table1 for ref-1