Part:BBa_K1796015
complete line of nif cluster
whole line of nif cluster,contain Pnif,nifBHDKENXV,hesA
Sequence and Features
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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).
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).
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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