Coding
Endolysin

Part:BBa_K2978000

Designed by: Jose Pablo Delgado Navarro   Group: iGEM19_Costa_Rica   (2019-10-09)
Revision as of 18:01, 20 October 2019 by PabloD CR (Talk | contribs)


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

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.


Characterization

T--Costa_Rica--team-R1.png

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]


[edit]
Categories
Parameters
device_type
positive_regulators
protein