Difference between revisions of "Part:BBa K907000"
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[[Image: KAIST_DNA_recombination_via_Bxb1_Int,_Xis.png |center|450px]]
 
[[Image: KAIST_DNA_recombination_via_Bxb1_Int,_Xis.png |center|450px]]
 
This part is a protein coding part which encodes Bxb1 gp35, which is a '''DNA integrase''' of ''Mycobacterium phage Bxb1''.
 
This part is a protein coding part which encodes Bxb1 gp35, which is a '''DNA integrase''' of ''Mycobacterium phage Bxb1''.
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The Bxb1 integrase is a DNA recombinase, more precisely a member of serine integrase family. It recognizes specific sequences, called ''attB'' and ''attP'', and then integrates, inverts, or excises dsDNA depending on the orientation of recognition sequences.
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<br>The Bxb1 integrase is a DNA recombinase, more precisely a member of serine integrase family. It recognizes specific sequences, called ''attB'' and ''attP'', and then integrates, inverts, or excises dsDNA depending on the orientation of recognition sequences.
 
We used this integrase to invert specific sequence in plasmid. The protein is well expressed in ''E.coli(strain MG1655)''
 
We used this integrase to invert specific sequence in plasmid. The protein is well expressed in ''E.coli(strain MG1655)''
 
When it inverts DNA sequence, the ''attB'' and ''attP'' sequences are changed into'' attL'' and ''attR'', as other DNA recombinases do. Another protein called Bxb1 gp47([https://parts.igem.org/Part:BBa_K907002 BBa_K0907002]) binds to integrase-DNA complexes and this complexes flips inverted DNA back into original sequence by regenerating ''attB'' and ''attP'' sequences.  
 
When it inverts DNA sequence, the ''attB'' and ''attP'' sequences are changed into'' attL'' and ''attR'', as other DNA recombinases do. Another protein called Bxb1 gp47([https://parts.igem.org/Part:BBa_K907002 BBa_K0907002]) binds to integrase-DNA complexes and this complexes flips inverted DNA back into original sequence by regenerating ''attB'' and ''attP'' sequences.  

Revision as of 18:12, 26 September 2012

Mycobacterium Phage Bxb1 gp35, DNA integrase


KAIST DNA recombination via Bxb1 Int, Xis.png

This part is a protein coding part which encodes Bxb1 gp35, which is a DNA integrase of Mycobacterium phage Bxb1.
The Bxb1 integrase is a DNA recombinase, more precisely a member of serine integrase family. It recognizes specific sequences, called attB and attP, and then integrates, inverts, or excises dsDNA depending on the orientation of recognition sequences. We used this integrase to invert specific sequence in plasmid. The protein is well expressed in E.coli(strain MG1655) When it inverts DNA sequence, the attB and attP sequences are changed into attL and attR, as other DNA recombinases do. Another protein called Bxb1 gp47(BBa_K0907002) binds to integrase-DNA complexes and this complexes flips inverted DNA back into original sequence by regenerating attB and attP sequences.

<Related Parts>
BBa_K907001 - Mycobacterium Phage Bxb1 excisionase
BBa_K907002 - Binary Signal Generator, RBS(reverse) - attB - Promoter - attP - RBS
BBa_K907003 - Binary Signal Generator, Promoter Reversed, RBS(reverse) - attB - Promoter(reverse) - attP - RBS
BBa_K907004 - Dual Phase Protein Generator(GFP default). mRFP and GFP
BBa_K907005 - Dual Phase Protein Generator(mRFP default). mRFP and GFP

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 192
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 466
    Illegal XhoI site found at 553
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1105
    Illegal NgoMIV site found at 1192
    Illegal AgeI site found at 242
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1300