Part:BBa_K2350018

J23106-B0034-Endolysin-B0010-B0012

Endolysin

　　Endolysin, also known as lysin or murein hydrolase, is peptidoglycan-hydrolyzing enzymes used by bacteriophages to degrade the bacterial host’s cell wall and lead to cell lysis. The gene sequences of endolysin in different species of bacteriophages are also slightly different. Most endolysin are monomeric proteins and consist of two types of domains, enzymatically active domains (EADs) and cell-binding domains (CBDs). EAD cleaves peptidoglycan bond, and CBD binds to the host’s cell wall. However, endolysin can’t pass through cytoplasmic membrane to access the cell wall alone. The lysis process includes another protein called holin, which can forms holes on cell membrane.

Promoter, RBS, and Terminator

　　 The endolysin in this part is from iGEM released part, BBa_K112806, which is endolysin from enterobacteria phage T4. Besides endolysin, in this composite part, we choose BBa_J23106 as a constitutive promoter, BBa_B0034 as ribosome binding site, BBa_B0010 and BBa_B0012 as double terminator, all of which are widely used parts in iGEM.

Usage and Biology

In 2017 NYMU project, we combine endolysin with constitutive promoter and holin with inducible promoter together, which is a regulatory suicide mechanism design. You can see more information in BBa_2350019 for holin part, and BBa_2350020 for part containing both endolysin and holin.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 334
  Illegal AgeI site found at 404
  
• 1000
  COMPATIBLE WITH RFC[1000]