Part:BBa_K2722002
6 times Chi recombination hotspot to inhibit Exonuclease V (RecBCD) activity
Chi is a short DNA Sequence, that is recognised by the RecBCD complex, also called Exonuclease V. Six Chi sites in a row can be used to inhibit RecBCD enzyme.
The RecBCD enzyme is both a helicase that unwinds, or separates the strands of DNA, and a nuclease that makes single-stranded nicks in DNA.(1) RecBCD complex is essential for cell viability as it plays an important role in repair of DNA double-strand breaks by initiating homologous recombination.(2)
However, RecBCD enzyme is also a highly processive at destruction of foreign DNA (3) The RecBCD enzyme is regulated by a cis-acting DNA sequence known as Chi (crossover hotspot instigator) that activates its recombination-promoting functions. Interaction with Chi causes an attenuation of the RecBCD enzyme's vigorous nuclease activity. (4)
Since Chi binds RecBCD with a greater affinity than other dsDNA substrate, this can be used to inhibit RecBCD activity in TX-TL, to more efficiently express linear DNA in TX-TL.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
//dna
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