Difference between revisions of "Part:BBa K4274009"
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sfp gene is a biobrick part from <i>B. amyloliquefaciens</i> FZB42 encoding 4'-phosphopantetheinyl transferase which functions as a primer of nonribosomal peptide synthesis via phosphopantetheinylation of thiotemplates. However, compared to the sfp gene in <i>Bacillus subtilis</i> 168, they only had an amino acid homology of 70%. And the mutation existed in <i>Bacillus subtilis</i> 168 resulted in its unable to produce any lipopeptide biosurfactant. Hence, to allow <i>B. subtilis</i> to produce fengycins, it is essential to first knock-out the mutant gene and then knock-in the correct sfp (Part: BBa_K4274009) and degQ (Part: BBa_K4274010).  
 
sfp gene is a biobrick part from <i>B. amyloliquefaciens</i> FZB42 encoding 4'-phosphopantetheinyl transferase which functions as a primer of nonribosomal peptide synthesis via phosphopantetheinylation of thiotemplates. However, compared to the sfp gene in <i>Bacillus subtilis</i> 168, they only had an amino acid homology of 70%. And the mutation existed in <i>Bacillus subtilis</i> 168 resulted in its unable to produce any lipopeptide biosurfactant. Hence, to allow <i>B. subtilis</i> to produce fengycins, it is essential to first knock-out the mutant gene and then knock-in the correct sfp (Part: BBa_K4274009) and degQ (Part: BBa_K4274010).  
  
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===Usage and Biology===
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==Usage and Biology==
 
Our sfp gene (Gene ID: 45022253) is a biobrick part derived from <i>B. amyloliquefaciens</i> FZB42 encoding 4'-phosphopantetheinyl transferase. As mentioned above, the correct sfp gene is the core of  lipopeptide biosurfactant production, thus the mutant sfp gene in <i>B. subtilis</i> is invalid.
 
Our sfp gene (Gene ID: 45022253) is a biobrick part derived from <i>B. amyloliquefaciens</i> FZB42 encoding 4'-phosphopantetheinyl transferase. As mentioned above, the correct sfp gene is the core of  lipopeptide biosurfactant production, thus the mutant sfp gene in <i>B. subtilis</i> is invalid.
 
<br>To construct a strain of <i>B. subtilis</i> to produce fengycins, we will use sfp_target (gRNA) (part: BBa_K4274008) to target the region of mutant sfp in <i>B. subtilis</i> for knock-out, simultaneously sfp (Part: BBa_K4274009) and degQ (Part: BBa_K4274010) gene were knocked-in in situ. It was used in the composite part PvanP*-sfp_target-sfp_HA_US-p43-K4274013-sfp-K4274014-degQ-sfp_HA_DS (Part: BBa_K4274035) to realize fengycins’ production in <i>B. subtilis</i>.
 
<br>To construct a strain of <i>B. subtilis</i> to produce fengycins, we will use sfp_target (gRNA) (part: BBa_K4274008) to target the region of mutant sfp in <i>B. subtilis</i> for knock-out, simultaneously sfp (Part: BBa_K4274009) and degQ (Part: BBa_K4274010) gene were knocked-in in situ. It was used in the composite part PvanP*-sfp_target-sfp_HA_US-p43-K4274013-sfp-K4274014-degQ-sfp_HA_DS (Part: BBa_K4274035) to realize fengycins’ production in <i>B. subtilis</i>.
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==Sequence and Features==
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<i>B. amyloliquefaciens</i> FZB42
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4274009 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4274009 SequenceAndFeatures</partinfo>
  
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K4274009 parameters</partinfo>
 
<partinfo>BBa_K4274009 parameters</partinfo>
 
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Revision as of 05:06, 12 October 2022

sfp

sfp gene is a biobrick part from B. amyloliquefaciens FZB42 encoding 4'-phosphopantetheinyl transferase which functions as a primer of nonribosomal peptide synthesis via phosphopantetheinylation of thiotemplates. However, compared to the sfp gene in Bacillus subtilis 168, they only had an amino acid homology of 70%. And the mutation existed in Bacillus subtilis 168 resulted in its unable to produce any lipopeptide biosurfactant. Hence, to allow B. subtilis to produce fengycins, it is essential to first knock-out the mutant gene and then knock-in the correct sfp (Part: BBa_K4274009) and degQ (Part: BBa_K4274010).

Usage and Biology

Our sfp gene (Gene ID: 45022253) is a biobrick part derived from B. amyloliquefaciens FZB42 encoding 4'-phosphopantetheinyl transferase. As mentioned above, the correct sfp gene is the core of lipopeptide biosurfactant production, thus the mutant sfp gene in B. subtilis is invalid.
To construct a strain of B. subtilis to produce fengycins, we will use sfp_target (gRNA) (part: BBa_K4274008) to target the region of mutant sfp in B. subtilis for knock-out, simultaneously sfp (Part: BBa_K4274009) and degQ (Part: BBa_K4274010) gene were knocked-in in situ. It was used in the composite part PvanP*-sfp_target-sfp_HA_US-p43-K4274013-sfp-K4274014-degQ-sfp_HA_DS (Part: BBa_K4274035) to realize fengycins’ production in B. subtilis.

Sequence and Features

B. amyloliquefaciens FZB42


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 38
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 38
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 38
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 38
    Illegal NgoMIV site found at 95
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 661

Functional Parameters