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 | + | 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 (Wang, <i>et al.</i>; 2015). |
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | + | 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. | |
| − | 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 | + | |
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Revision as of 05:23, 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 (Wang, et al.; 2015).
Usage and Biology
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]