Generator

Part:BBa_K1934060

Designed by: Mathilde Gonthier   Group: iGEM16_INSA-Lyon   (2016-10-13)
Revision as of 14:34, 15 October 2016 by MattBorel (Talk | contribs)


p51 subunit of HIV reverse transcriptase

This part contains the sequence of the p51 subunit of HIV reverse transcriptase. It is not functional on his own, to have an activity it must be associated with the p66 protein.

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 1100
    Illegal BglII site found at 1150
    Illegal XhoI site found at 1419
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    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. Purifying the part

This part was cloned into e. coli NM522. Then the bacterium were grown in LB overnight at 37°. We got an OD600 of 2.4 at this stage, enough for harvest. The p51 has been tagged with a polyHis tag. A NiNTA column is used to purify it. We used NiNTA columns kit from kiagen. The protocol may be downloaded on this page. The protein purification yield was 350% and the global recovery yield was 24 %.

Figure 1. Steps of purification of the p51 /b> Lane A shows the raw cellular extract. Lanes B,C,D and E shows the different wash step that were performed. On these lanes the p51 is not visible. Lanes F and G shows the purified recovered protein.

1. Purifying the part

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