Difference between revisions of "Part:BBa K1934061"
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This part contains the sequence of the p66 subunit of HIV reverse transcriptase.
 
This part contains the sequence of the p66 subunit of HIV reverse transcriptase.
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This part has been transferred into e. coli NM522 for expression. After a 6 hours pre culture phase at 37° , the expression culture was grown overnight at 30°. We the used instructions provided on the NiNTA columns kit from Quigen to purify the protein. We obtained
 
  
 
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<partinfo>BBa_K1934061 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1934061 SequenceAndFeatures</partinfo>
  
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=='''Characterization'''==
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<html>
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<h3 id="degradation0301">1. Testing the activity of the part </h3>
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<p>This part has been transferred into e. coli NM522 for expression. After a 6 hours pre culture phase at 37° , the expression culture was grown overnight at 30°. We the used instructions provided on the NiNTA columns kit from Quigen to purify the protein. We obtained </p>
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<figure><img src="https://static.igem.org/mediawiki/2014/f/f8/Peking2014jyj_3.png"/><figcaption><b>Figure 2. Measurement of MlrA activity.</b> The OD405 indicates the concentration of pNP, and the change of pNP level could reflect the PP1 activity(a). MC can strongly inhibit the PP1 activity(b), and the MlrA can cleave the MC and dampen its toxicity(c).</figcaption></figure>
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</html>
  
 
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<!-- Uncomment this to enable Functional Parameter display  

Revision as of 13:59, 15 October 2016


p66 subunit of HIV reverse transcriptase

This part contains the sequence of the p66 subunit of HIV reverse transcriptase.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 238
    Illegal BglII site found at 937
    Illegal BglII site found at 1100
    Illegal BglII site found at 1150
    Illegal XhoI site found at 1800
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1739
    Illegal AgeI site found at 1067
    Illegal AgeI site found at 1182
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 20

Characterization

1. Testing the activity of the part

This part has been transferred into e. coli NM522 for expression. After a 6 hours pre culture phase at 37° , the expression culture was grown overnight at 30°. We the used instructions provided on the NiNTA columns kit from Quigen to purify the protein. We obtained

Figure 2. Measurement of MlrA activity. The OD405 indicates the concentration of pNP, and the change of pNP level could reflect the PP1 activity(a). MC can strongly inhibit the PP1 activity(b), and the MlrA can cleave the MC and dampen its toxicity(c).