Difference between revisions of "Part:BBa K2978000"

 
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This part makes references to an endolysin from a bacteriophage that infects <i>Clostridium difficile</i>. It is section of the complete protein CD27L, which in the residues 2 to 143 has its domain homologous to <i>N-acetyl-muramoyl-L-alanine</i>  amidases. In this part we synthesize  de novo the residues 1 to 179 of the protein, with codon optimization for <i>E.coli</i>. In previously works this reduced protein has shows a faster lysis to different strains of the pathogen and a high level of selectivity against clostridia [1].
 
This part makes references to an endolysin from a bacteriophage that infects <i>Clostridium difficile</i>. It is section of the complete protein CD27L, which in the residues 2 to 143 has its domain homologous to <i>N-acetyl-muramoyl-L-alanine</i>  amidases. In this part we synthesize  de novo the residues 1 to 179 of the protein, with codon optimization for <i>E.coli</i>. In previously works this reduced protein has shows a faster lysis to different strains of the pathogen and a high level of selectivity against clostridia [1].
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The expression of this parts is under a T7 promoter, this mean that if we want to induce this protein we have to use a chassis with T7. In our case we confirm that <i>E. coli</i> BL21 de3 produce the lysin when IPTG is applied, because in this strain the polymerase is under a pLac promoter (Figure 1).
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The expression of this parts is under a T7 promoter, this mean that if we want to induce this protein we have to use a chassis with T7. In our case we confirm that <i>E. coli</i> BL21 de3 produce the lysin when IPTG is applied, because in this strain the polymerase is under a pLac promoter.
  
  

Revision as of 05:06, 10 October 2019


Causes cell lysis of the pathogen Clostridium difficile

This part makes references to an endolysin from a bacteriophage that infects Clostridium difficile. It is section of the complete protein CD27L, which in the residues 2 to 143 has its domain homologous to N-acetyl-muramoyl-L-alanine amidases. In this part we synthesize de novo the residues 1 to 179 of the protein, with codon optimization for E.coli. In previously works this reduced protein has shows a faster lysis to different strains of the pathogen and a high level of selectivity against clostridia [1]. The expression of this parts is under a T7 promoter, this mean that if we want to induce this protein we have to use a chassis with T7. In our case we confirm that E. coli BL21 de3 produce the lysin when IPTG is applied, because in this strain the polymerase is under a pLac promoter.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]