Difference between revisions of "Part:BBa K2978000"

Revision as of 18:03, 10 October 2019

Causes cell lysis of the pathogen Clostridium difficile

This part makes reference to an endolysin from a bacteriophage that infects Clostridium difficile. It is a segment of the complete protein CD27L, which residues from 2 to 143 are homolougous to the domain N-acetyl-muramoyl-L-alanine amidase. This part codes for residues 1 to 179 of the mentioned protein. It has been codon optimized for E.coli. In previous works this domain has shown a faster lysis to different strains of the pathogen and a high level of selectivity against clostridia compared to the complete protein (Wang, et al., 2015).

Usage and Biology

The expression of this part is under a T7 promoter, therefore the chasis must contain the T7 polymerase. In our lab we confirmed that E. coli BL21 de3 produces the lysin when IPTG is applied, as this strain has the respective polymerase under 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]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2978000 short</partinfo>
-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).
+This part makes reference to an endolysin from a bacteriophage that infects <i>Clostridium difficile</i>. It is a segment of the complete protein CD27L, which residues from 2 to 143 are homolougous to the domain <i>N-acetyl-muramoyl-L-alanine</i> amidase. This part codes for residues 1 to 179 of the mentioned protein. It has been codon optimized for <i>E.coli</i>. In previous works this domain has shown a faster lysis to different strains of the pathogen and a high level of selectivity against clostridia compared to the complete protein (Wang, <i>et al.</i>, 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 <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 part is under a T7 promoter, therefore the chasis must contain the T7 polymerase. In our lab we confirmed that <i>E. coli</i> BL21 de3 produces the lysin when IPTG is applied, as this strain has the respective polymerase under pLac promoter.
 <!-- -->