Difference between revisions of "Part:BBa K1934061"
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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 could effectively purify the protein, with a purification yield of 350% and a global yield of 24 % </p>
 
<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 could effectively purify the protein, with a purification yield of 350% and a global yield of 24 % </p>
  
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<figure><img src="https://static.igem.org/mediawiki/2016/e/e9/INSA-Lyon_p51.png" width = "400" /><figcaption><b>Figure 1. Purification of the p51.</b> Lane A show the raw cellular extract. Lanes F and G contains the purified protein after some washes. Lanes B,C,D and E represents the different wha steps, the p51 is not present in these lanes. </figcaption></figure>
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<figure><img src="https://static.igem.org/mediawiki/2016/e/e9/INSA-Lyon_p51.png" width = "400" align="middle" /><figcaption><b>Figure 1. Purification of the p51.</b> Lane A show the raw cellular extract. Lanes F and G contains the purified protein after some washes. Lanes B,C,D and E represents the different wha steps, the p51 is not present in these lanes. </figcaption></figure>
  
  

Revision as of 14:11, 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 could effectively purify the protein, with a purification yield of 350% and a global yield of 24 %

Figure 1. Purification of the p51. Lane A show the raw cellular extract. Lanes F and G contains the purified protein after some washes. Lanes B,C,D and E represents the different wha steps, the p51 is not present in these lanes.