User:Pakpoom Subsoontorn/Notebook/Genetically Encoded Memory/2008/10/26

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Profile

 * PDB ID:
 * Source: Lambda phage's enzyme functioning in E.coli host
 * Classification Keyword: integrase, tyrosine recombinase
 * Size: 356 aa
 * Enzyme Structure
 * crystal structure of C-terminal catalytic domain is known
 * domains
 * N-terminal 1-64 aa binds to arm-type site.
 * C-terminal 65-169 aa binds to low affinity core-type site.
 * 170-356 aa is the minimal catalytic domain which include six tyrosine kinase conserved residues: R212, K235, H308, R311, Y342, H308 (nucleophilic tyrosine and catalytic pentad)
 * Att-site
 * attB:: has two inverted repeated sequence flanking an overlap region. Inverted repeated sequence (core-type) is about 9-13 bps.
 * attP is more complicated, with arm-type sites flanking core-type site.
 * The overlap region (between attP and attB) is about 6-8 bps and shares sequence with attP. Here is where crossing over occur. Overlap-region:TTTATAC:
 * auxiliary factors:
 * Mechanism: high affinity binding at arm-region of attP, low affinity binding at core-region. Integration host factor (IHF, from HimA and hip genes) is required for sharp bending in DNA, allowing the integrase bound at arm-type site to also bind at the low affinity core-type sites. Break one strand at the time. Tyrosine phosphate binds to 3'end . 5'OH is free. Integration involve holiday junction
 * Engineering
 * Invitrogen Gateway (r) system

Reference

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