Difference between revisions of "Part:BBa K1797013"

Latest revision as of 15:30, 18 September 2015

Exo-an exonuclease in the Red homologous recombination system

Fig.1 The first step of gene editing using red recombination system (source: http://www.sci.sdsu.edu/~smaloy/MicrobialGenetics/topics/in-vitro-genetics/red-swap.html)

Fig.2 The second step of gene editing using red recombination system (source: http://www.sci.sdsu.edu/~smaloy/MicrobialGenetics/topics/in-vitro-genetics/red-swap.html)

Red recombination system is a efficient homologous recombination system found in lamda phage. It includes three main proteins, gam, bet and exo. Gam is able to bind RecBCD, inhibiting its ability to degrade exogenous linear DNA. Bet binds to the 3' overhang produced by exo and protects it, preventing its degradation by nucleases. Red recombination system is becoming popular in terms of gene editing. The common procedure is to PCR a certain fragment(often a kana cassette for easy screening later) flanked with homologous regions(about 20-50bp) first. Then the linear fragment and the red recombination system is introduced into bacteria for homologous recombination.

This part contains one component of red recombination system-exo. Exo is an exonuclease which can bite a dsDNA and form a 3' overhang.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 199
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1797013 short</partinfo>
-[[File:Red-pcr.gif|400px|thumb|left|Fig.1 The first step of gene editing using red recombination system (source: http://www.sci.sdsu.edu/~smaloy/MicrobialGenetics/topics/in-vitro-genetics/red-swap.html)]]
+[[File:Red-pcr.gif|400px|thumb|center|Fig.1 The first step of gene editing using red recombination system (source: http://www.sci.sdsu.edu/~smaloy/MicrobialGenetics/topics/in-vitro-genetics/red-swap.html)]]
-[[File:Red-rec.gif‎|400px|thumb|left|Fig.2 The second step of gene editing using red recombination system (source: http://www.sci.sdsu.edu/~smaloy/MicrobialGenetics/topics/in-vitro-genetics/red-swap.html)]]
+[[File:Red-rec.gif‎|400px|thumb|center|Fig.2 The second step of gene editing using red recombination system (source: http://www.sci.sdsu.edu/~smaloy/MicrobialGenetics/topics/in-vitro-genetics/red-swap.html)]]
 Red recombination system is a efficient homologous recombination system found in lamda phage. It includes three main proteins, gam, bet and exo. Gam is able to bind RecBCD, inhibiting its ability to degrade exogenous linear DNA. Bet binds to the 3' overhang produced by exo and protects it, preventing its degradation by nucleases. Red recombination system is becoming popular in terms of gene editing. The common procedure is to PCR a certain fragment(often a kana cassette for easy screening later) flanked with homologous regions(about 20-50bp) first. Then the linear fragment and the red recombination system is introduced into bacteria for homologous recombination.