Difference between revisions of "Part:BBa K1155007"
 
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lacZ gene encodes the protein beta-galactosidase which causes bacteria expressing the gene to appear blue when grown on a medium that contains the substrate analog X-gal.
 
lacZ gene encodes the protein beta-galactosidase which causes bacteria expressing the gene to appear blue when grown on a medium that contains the substrate analog X-gal.
  
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In order to be easily used by all the other teams, our team improved the biobrick BBa_I732017 designed by iGEM07_USTC by asembly of biobrick BBa_I732017 and B0015.
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In order to be easily used by all the other teams, our team improved the biobrick [https://parts.igem.org/Part:BBa_I732017 BBa_I732017] designed by iGEM07_USTC by asembly of biobrick BBa_I732017 and B0017.
  
 
A composite part constituted of a RBS, full length lacZ gene and a terminator. We used it as one of our reporter gene in ''Escherichia coli''.  
 
A composite part constituted of a RBS, full length lacZ gene and a terminator. We used it as one of our reporter gene in ''Escherichia coli''.  
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<partinfo>BBa_K1155007 parameters</partinfo>
 
<partinfo>BBa_K1155007 parameters</partinfo>
 
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'''Characterization'''
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* We have tested the Biobricks BBa_K1155007 in the construction: BBa_K1155004 + BBa_K1155007 in pSB1C3
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BBa_K1155004 is a promoteur PnirB activated in anaerobic condition by binding of the FNR regulator (see BBa_K1155004 for more information).
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What we expected :
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+ O2 : FNR inactivated : PnirB inactivated --> lacZ not expressied --> no color
 +
 +
- O2 : FNR activated : PnirB activated --> lacZ expressed --> blue color
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What we observed :
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We observe  blue color in both conditions. This must be because we introduce our construction (BBa_K1155004+BBa_K1155007) in a high copy backbone.
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[[File:Ps004.jpg|500px]]
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* We have tested the Biobricks BBa_K1155007 in the construction: BBa_K1155006 + BBa_K1155007 in pSB1K3
 +
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BBa_K1155006 is a promoteur PnarK activated in anaerobic condition by binding of the FNR regulator (see BBa_K1155006 for more information).
 +
 +
What we expected :
 +
 +
+ O2 : FNR inactivated : PnirB inactivated --> lacZ not expressied --> no color
 +
 +
- O2 : FNR activated : PnirB activated --> lacZ expressed --> blue color
 +
 +
What we observed :
 +
 +
We observe  blue color in anaerobic conditions. We could not do this experience in aerobic conditions.
 +
 +
[[File:Ps007.jpg|500px]]
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 +
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<b> TO CONCLUDE : Our biobrick BBa_K1155007 works very well because we can observe blue colonies on Xgal. It can be easily used as promoter expression rapporter </b>

Latest revision as of 07:58, 5 October 2013

RBS-lacZ-term


lacZ gene encodes the protein beta-galactosidase which causes bacteria expressing the gene to appear blue when grown on a medium that contains the substrate analog X-gal.

In order to be easily used by all the other teams, our team improved the biobrick BBa_I732017 designed by iGEM07_USTC by asembly of biobrick BBa_I732017 and B0017.

A composite part constituted of a RBS, full length lacZ gene and a terminator. We used it as one of our reporter gene in Escherichia coli.


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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization

  • We have tested the Biobricks BBa_K1155007 in the construction: BBa_K1155004 + BBa_K1155007 in pSB1C3

BBa_K1155004 is a promoteur PnirB activated in anaerobic condition by binding of the FNR regulator (see BBa_K1155004 for more information).

What we expected :

+ O2 : FNR inactivated : PnirB inactivated --> lacZ not expressied --> no color

- O2 : FNR activated : PnirB activated --> lacZ expressed --> blue color

What we observed :

We observe blue color in both conditions. This must be because we introduce our construction (BBa_K1155004+BBa_K1155007) in a high copy backbone.

Ps004.jpg

  • We have tested the Biobricks BBa_K1155007 in the construction: BBa_K1155006 + BBa_K1155007 in pSB1K3

BBa_K1155006 is a promoteur PnarK activated in anaerobic condition by binding of the FNR regulator (see BBa_K1155006 for more information).

What we expected :

+ O2 : FNR inactivated : PnirB inactivated --> lacZ not expressied --> no color

- O2 : FNR activated : PnirB activated --> lacZ expressed --> blue color

What we observed :

We observe blue color in anaerobic conditions. We could not do this experience in aerobic conditions.

Ps007.jpg


TO CONCLUDE : Our biobrick BBa_K1155007 works very well because we can observe blue colonies on Xgal. It can be easily used as promoter expression rapporter