Difference between revisions of "Part:BBa K2374001:Design"

(→‎Design Notes)
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
  
−
We get this 452bp sequence from FlyBase ([http://flybase.org/reports/FBgn0005626.html] Dmel\ple)
+
We get this 452bp sequence from FlyBase ( Dmel\ple [http://flybase.org/reports/FBgn0005626.html]). We regard the 5'-UTR region as promoter region of ''ple''.
 +
This should not be the most accurate and concise promoter sequence. We hope that subsequent users will be able to accurately identify the promoter sequence.
 +
We cloned this 452bp TH promoter easily from ''D. melanogaster'' 's cDNA library and the sequencing result is correct.
 +
We combined this promoter with UAS/GAL4 system to achieve gene expression in specific cells.
 +
We connected TH promoter to GAL4( BBa_K2374004 [https://parts.igem.org/Part:BBa_K2374004] )and GAL80ts( BBa_K2374002 [https://parts.igem.org/Part:BBa_K2374002] )respectively, and cloned them into pUAST vector which had been removed the UAS sequence( BBa_K2374008 [https://parts.igem.org/Part:BBa_K2374008] )
  
  

Revision as of 03:17, 30 October 2017


TH (ple) promoter-> (fruit fly)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 137
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We get this 452bp sequence from FlyBase ( Dmel\ple [http://flybase.org/reports/FBgn0005626.html]). We regard the 5'-UTR region as promoter region of ple. This should not be the most accurate and concise promoter sequence. We hope that subsequent users will be able to accurately identify the promoter sequence. We cloned this 452bp TH promoter easily from D. melanogaster 's cDNA library and the sequencing result is correct. We combined this promoter with UAS/GAL4 system to achieve gene expression in specific cells. We connected TH promoter to GAL4( BBa_K2374004 [1] )and GAL80ts( BBa_K2374002 [2] )respectively, and cloned them into pUAST vector which had been removed the UAS sequence( BBa_K2374008 [3] )


site direct mutangenesis: EcoR I (184) GAATTC->GATTTC Xba I (219) TCTAGA->TGTAGA

Source

NT_037436.4 (NCBI)

References