Part:BBa_K2350019
R0010-B0034-Holin-B0010-B0012
Holin
Holin is a group of membrane protiens produced by bacteriophages. Holin can forms holes on the bacteria host’s cell membrane and control the access of endolysin to peptidoglycan in cell wall. Endolysin is peptidoglycan-hydrolyzing enzymes and is able to degrade the bacteria host’s cell wall, leading to cell lysis. Therefore, the expression and activation of holin determines the cell lysis at a precise time.
T4 Holin
The gene sequences and the structures of holin in different species of bacteriophages are also different. The holin in this part is T4 holin. T4 holin consists of one transmembrane domain, C-terminus in the periplasm, and N-terminus in the cytoplasm. The holin in this part is from iGEM released parts, BBa_K112000, which is holin from enterobacteria phage T4.
Promoter, RBS, and Terminator
In our 2017 NYMU project, we combine T4 holin and T4 endolysin in a construct to make a E.coli suicide part. (You can see more information in BBa_2350018 for endolysin part, and BBa_2350020 for part containing both endolysin and holin.) To control the precise suicide timing, we use an inducible promoter for holin. We choose a lactose-induced promoter, BBa_R0010, to regulate this suicide mechanism. Besides holin and inducible promoter, in this composite part, we choose 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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