Composite

Part:BBa_K1796015

Designed by: Nannan Xie   Group: iGEM15_SCU_China   (2015-09-18)
Revision as of 11:08, 15 October 2018 by Manbu (Talk | contribs)


complete line of nif cluster

whole line of nif cluster,contain Pnif,nifBHDKENXV,hesA

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2163
    Illegal EcoRI site found at 6951
    Illegal PstI site found at 2689
    Illegal PstI site found at 6597
    Illegal PstI site found at 7295
    Illegal PstI site found at 7866
    Illegal PstI site found at 7937
    Illegal PstI site found at 8743
    Illegal PstI site found at 8983
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2163
    Illegal EcoRI site found at 6951
    Illegal PstI site found at 2689
    Illegal PstI site found at 6597
    Illegal PstI site found at 7295
    Illegal PstI site found at 7866
    Illegal PstI site found at 7937
    Illegal PstI site found at 8743
    Illegal PstI site found at 8983
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2163
    Illegal EcoRI site found at 6951
    Illegal BglII site found at 3655
    Illegal BglII site found at 10747
    Illegal BamHI site found at 10570
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2163
    Illegal EcoRI site found at 6951
    Illegal PstI site found at 2689
    Illegal PstI site found at 6597
    Illegal PstI site found at 7295
    Illegal PstI site found at 7866
    Illegal PstI site found at 7937
    Illegal PstI site found at 8743
    Illegal PstI site found at 8983
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2163
    Illegal EcoRI site found at 6951
    Illegal PstI site found at 2689
    Illegal PstI site found at 6597
    Illegal PstI site found at 7295
    Illegal PstI site found at 7866
    Illegal PstI site found at 7937
    Illegal PstI site found at 8743
    Illegal PstI site found at 8983
    Illegal NgoMIV site found at 5942
    Illegal NgoMIV site found at 6189
    Illegal NgoMIV site found at 7624
    Illegal AgeI site found at 1136
    Illegal AgeI site found at 2096
    Illegal AgeI site found at 2451
    Illegal AgeI site found at 3680
    Illegal AgeI site found at 4362
    Illegal AgeI site found at 4735
    Illegal AgeI site found at 5202
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2268


iGEM2018_Nanjing China Experiment

This year our team use the nitrogen fixation gene cluster from Paenibacillus polymyxa CR1, which shares a close biological relationship with Paenibacillus sp. WLY78, so the data can provide some reference to this part.
In order to test the expression efficiency of the nif cluster,firstly we measured the transcriptional activity of nif promoter by combining it with the gene of fluorescent protein Dronpa, and use T5 (IPTG Inducible) Promoter, BBa_M50075 as a positive control(Fig 1).

Fig 2：Expression efficiency of Pnif

Comparison of the expression efficiency of Pnif and T5 (IPTG Inducible) Promoter.
T5 (IPTG Inducible) Promoter BBa_M50075; Pnif: nif promoter BBa_K1796001.

As demonstrated above, nif promoter is quite strong,however, how capable it is in our nitrogen fixation system remains unclear. So we test expression profiles of each structure gene in the nif cluster that overexpressed in EJNC by conducting Real-time Quantitative PCR(qPCR). Relative expression compared to the housekeeping gene 16S rRNA is shown(Fig2).

Fig 2. The qPCR results for components of nitrogen fixation system

qRT-PCR analysis demonstrates that all the component genes of the nif cluster are significantly over expressed in EJNC whereas the transcription of these genes are no detected (N.D.) in nondiazotrophic E.coli JM109.

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