Difference between revisions of "Part:BBa K3467005"

 
 
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_NOTOC__
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<partinfo>BBa_K3467005 short</partinfo>
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3467005 SequenceAndFeatures</partinfo>
  
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We obtained PMaGPD promoter from Metarhizium acridum. Comparing with GPD promoter for fungi plasmid, obtained from Aspergillus nidulans, this promoter was analyzed by 5’ -deletion strategy with GUS gene as reporter gene. The analysis from the reference reveals that PMaGPD includes the 1.7 kb region upstream of the start codon of the Magpd gene. According to the experiment, deletion from the -1691bp to -1463bp didn’t significantly affect the PMagpd activity. We obtained Pmagpd sequence by using PCR and cloning from the extract Metarhizium acridum’s genome. Here is our electrophoresis picture of PMagpd with 1463bp.
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[[File:T--CAU_China--PMagpd_promoter.jpg|500px|thumb|center|'''Fig. 1''' PMagpd promoter]]
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Comparing to the transcriptional ability from the reference, PMagpd has a relative activity 1.4 times higher than gpd promoter from Aspergillus nidulans. As a result, we decided to use PMagpd as our Metarhizium acridum promoter basic part.

Latest revision as of 02:35, 27 October 2020

_NOTOC__ Promoter of Metarhizium acridums GPD Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 551
    Illegal EcoRI site found at 1128
    Illegal SpeI site found at 673
    Illegal PstI site found at 344
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 551
    Illegal EcoRI site found at 1128
    Illegal SpeI site found at 673
    Illegal PstI site found at 344
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 551
    Illegal EcoRI site found at 1128
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 551
    Illegal EcoRI site found at 1128
    Illegal SpeI site found at 673
    Illegal PstI site found at 344
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 551
    Illegal EcoRI site found at 1128
    Illegal SpeI site found at 673
    Illegal PstI site found at 344
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 245
    Illegal BsaI.rc site found at 1437

We obtained PMaGPD promoter from Metarhizium acridum. Comparing with GPD promoter for fungi plasmid, obtained from Aspergillus nidulans, this promoter was analyzed by 5’ -deletion strategy with GUS gene as reporter gene. The analysis from the reference reveals that PMaGPD includes the 1.7 kb region upstream of the start codon of the Magpd gene. According to the experiment, deletion from the -1691bp to -1463bp didn’t significantly affect the PMagpd activity. We obtained Pmagpd sequence by using PCR and cloning from the extract Metarhizium acridum’s genome. Here is our electrophoresis picture of PMagpd with 1463bp.

Fig. 1 PMagpd promoter

Comparing to the transcriptional ability from the reference, PMagpd has a relative activity 1.4 times higher than gpd promoter from Aspergillus nidulans. As a result, we decided to use PMagpd as our Metarhizium acridum promoter basic part.