Difference between revisions of "Part:BBa K3552006"

 
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__NOTOC__
 
__NOTOC__
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<partinfo>BBa_K3552006 short</partinfo>
 
<partinfo>BBa_K3552006 short</partinfo>
  
 
HofB is a transport protein found in bacteria and is ATP-binding and nucleotide-binding, constitute the generator of pilA. HofC is an inner membrane protein, a base for the generator of pilA. This part is in the part collection where we have 12 genes that code for the base generator of pilA.  
 
HofB is a transport protein found in bacteria and is ATP-binding and nucleotide-binding, constitute the generator of pilA. HofC is an inner membrane protein, a base for the generator of pilA. This part is in the part collection where we have 12 genes that code for the base generator of pilA.  
  
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The hofC is from the hof operon encoding the generator of major pilin, the ATPase HofB and the platform protein HofC is localized at 3 min of E.coli map. The HofB gene is a ATPase motor which localized at the base of the pilin generator system in the inner membrane, propel the assembly of pilus and it interacts with the platform protein HofC and HofM component of the inner membrane anchored assembly platform complex that also includes HofN and HofO. The hofMNOPQ operon encodes the assembly platform complex connecting HofB ATPase with the secretin hofQ.
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The part collection includes:
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Parts that are different kinds of type 4 pilus:
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<partinfo>BBa_K3552003</partinfo> 
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<partinfo>BBa_K3552004</partinfo>
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<partinfo>BBa_K3552005</partinfo>
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<partinfo>BBa_K3552006</partinfo>
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<partinfo>BBa_K3552007</partinfo>
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<partinfo>BBa_K3552008</partinfo>
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<partinfo>BBa_K3552018</partinfo> 
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<partinfo>BBa_K3552019</partinfo> 
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<partinfo>BBa_K3552020</partinfo> 
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<partinfo>BBa_K3552021</partinfo> 
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<partinfo>BBa_K3552022</partinfo> 
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<partinfo>BBa_K3552023</partinfo> 
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<partinfo>BBa_K3552024</partinfo> 
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<partinfo>BBa_K3552025</partinfo> 
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<partinfo>BBa_K3552026</partinfo> 
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<partinfo>BBa_K3552027</partinfo> 
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<partinfo>BBa_K3552028</partinfo> 
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<partinfo>BBa_K3552029</partinfo>  
  
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<!-- Add more about the biology of this part here
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Our part collection can instruct other teams to designed new rechargeable pilus and substitution of different major pilin.
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===Usage and Biology===
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<!-- -->
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<span class='h3bb'>'''Sequence and Features'''</span>
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3552006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3552006 SequenceAndFeatures</partinfo>
  
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==Usage and Biology==
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The hofC is from the hof operon encoding the generator of major pilin, the ATPase HofB and the platform protein HofC is localized at 3 min of E.coli map. The HofB gene is a ATPase motor which localized at the base of the pilin generator system in the inner membrane, propel the assembly of pilus and it interacts with the platform protein HofC and HofM component of the inner membrane anchored assembly platform complex that also includes HofN and HofO. The  hofMNOPQ operon encodes the assembly platform complex connecting HofB ATPase with the secretin hofQ.
 +
 +
==Characterization==
 +
We first uses the bacteria DH5a as the template and obtain the DNA fragment of 12 genes. Point mutations are carried out to alter the sequence into the same as the EHEC genes. The ygdB gene is reconstructed through the SOE pcr as it is so different from the required sequence. We verified the practicability of the generator through the western blot of pilA in vivo which confirmed the usage of the generator because the PilA is able to display outside the cell.
  
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<!-- Uncomment this to enable Functional Parameter display
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<partinfo>BBa_K3552000 parameters</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K3552006 parameters</partinfo>
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Revision as of 03:48, 26 October 2020


hofB-hofC

HofB is a transport protein found in bacteria and is ATP-binding and nucleotide-binding, constitute the generator of pilA. HofC is an inner membrane protein, a base for the generator of pilA. This part is in the part collection where we have 12 genes that code for the base generator of pilA.

The part collection includes: Parts that are different kinds of type 4 pilus: BBa_K3552003 BBa_K3552004 BBa_K3552005 BBa_K3552006 BBa_K3552007 BBa_K3552008 BBa_K3552018 BBa_K3552019 BBa_K3552020 BBa_K3552021 BBa_K3552022 BBa_K3552023 BBa_K3552024 BBa_K3552025 BBa_K3552026 BBa_K3552027 BBa_K3552028 BBa_K3552029

Our part collection can instruct other teams to designed new rechargeable pilus and substitution of different major pilin.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 872
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1801
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage and Biology

The hofC is from the hof operon encoding the generator of major pilin, the ATPase HofB and the platform protein HofC is localized at 3 min of E.coli map. The HofB gene is a ATPase motor which localized at the base of the pilin generator system in the inner membrane, propel the assembly of pilus and it interacts with the platform protein HofC and HofM component of the inner membrane anchored assembly platform complex that also includes HofN and HofO. The hofMNOPQ operon encodes the assembly platform complex connecting HofB ATPase with the secretin hofQ.

Characterization

We first uses the bacteria DH5a as the template and obtain the DNA fragment of 12 genes. Point mutations are carried out to alter the sequence into the same as the EHEC genes. The ygdB gene is reconstructed through the SOE pcr as it is so different from the required sequence. We verified the practicability of the generator through the western blot of pilA in vivo which confirmed the usage of the generator because the PilA is able to display outside the cell.