Difference between revisions of "Part:BBa K873003"
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K873003 short</partinfo>
 
<partinfo>BBa_K873003 short</partinfo>
 +
 
This part contains a reporter gene I13521,combined with OriTR(the R plasmid nic region, is where the relaxosome nicks the plasmid and conjugative transfer by R plasmid machinery begins.)   
 
This part contains a reporter gene I13521,combined with OriTR(the R plasmid nic region, is where the relaxosome nicks the plasmid and conjugative transfer by R plasmid machinery begins.)   
−
<!-- Add more about the biology of this part here
 
−
Once the conjugation take place,the recipient bacterium will obtain the reporter gene which gives us a signal to distinguish whether the conjugation has happened as well as the  conjugation efficiency easily though observation. In addition, it facilitates the Follow-up experiments on the conjugation rate, conjugation time, and conjugation efficiency between different strains.
 
  
 +
==Function ==
 +
 +
Once the conjugation take place,the recipient bacterium will obtain the reporter gene which gives
 +
us a signal to distinguish whether the conjugation has happened as well as the conjugation efficiency
 +
easily through observation. In addition, it facilitates the Follow-up experiments on the conjugation
 +
rate, conjugation time, and conjugation efficiency between different strains.
 +
 +
== Results ==
 +
The TEM image of conjugation:
 +
 +
[[Image:fig1.jpg]]
 +
 +
The agar Luria-Bertani medium of DH5α:  --------------------------------------------------------------    The agar Luria-Bertani medium of BL21:
 +
 +
[[Image:seulfig2.jpg]]                            [[Image:seulfig3.jpg]]   
 +
 +
 +
The agar Luria-Bertani medium of HB101:  --------------------------------------------------------------    The agar Luria-Bertani medium of conjugation:
 +
 +
[[Image:seulfig5.jpg]]                            [[Image:seulfig4.jpg]] 
 +
 +
It's show that the conjugation system did work.
 +
For more information on how this part operates in our system, please visit [http://2012.igem.org/Team:SEU_A SEU_A ]．
 +
 +
== Safety ==
 +
The biobrick safety matters that we may involved in is that we construct a plasmid with the conjugation
 +
function in order to spread our target DNA fragment. For the sake of preventing the plasmid broadcast some
 +
unknown function genes, we ensure that every used liquid medium and gel solid medium are sterile before
 +
disposal. Well, all the other DNA fragments no matter we used or modified are achieved from the kit plates
 +
or commonly employed in laboratory.
 +
 +
 +
== references ==
 +
         
 +
[http://onlinelibrary.wiley.com/doi/10.1002/bmb.113/full]
 +
 +
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
−
Once the conjugation take place,the recipient bacterium will obtain the reporter gene which gives us a signal to distinguish whether the conjugation has happened as well as the  conjugation efficiency easily though observation. In addition, it facilitates the Follow-up experiments on the conjugation rate, conjugation time, and conjugation efficiency between different strains.
 
 
<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 22:15, 26 September 2012

conjugative transfer of RFG

This part contains a reporter gene I13521,combined with OriTR(the R plasmid nic region, is where the relaxosome nicks the plasmid and conjugative transfer by R plasmid machinery begins.)

Function

Once the conjugation take place,the recipient bacterium will obtain the reporter gene which gives
us a signal to distinguish whether the conjugation has happened as well as the conjugation efficiency
easily through observation. In addition, it facilitates the Follow-up experiments on the conjugation
rate, conjugation time, and conjugation efficiency between different strains.

Results

The TEM image of conjugation:

Fig1.jpg

The agar Luria-Bertani medium of DH5α: -------------------------------------------------------------- The agar Luria-Bertani medium of BL21:

Seulfig2.jpg Seulfig3.jpg


The agar Luria-Bertani medium of HB101: -------------------------------------------------------------- The agar Luria-Bertani medium of conjugation:

Seulfig5.jpg Seulfig4.jpg 

It's show that the conjugation system did work.
For more information on how this part operates in our system, please visit [http://2012.igem.org/Team:SEU_A SEU_A ]．

Safety

The biobrick safety matters that we may involved in is that we construct a plasmid with the conjugation
function in order to spread our target DNA fragment. For the sake of preventing the plasmid broadcast some
unknown function genes, we ensure that every used liquid medium and gel solid medium are sterile before 
disposal. Well, all the other DNA fragments no matter we used or modified are achieved from the kit plates
or commonly employed in laboratory.


references

[http://onlinelibrary.wiley.com/doi/10.1002/bmb.113/full]

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 NgoMIV site found at 117
    Illegal NgoMIV site found at 127
    Illegal AgeI site found at 1014
    Illegal AgeI site found at 1126
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 19