Difference between revisions of "Part:BBa K3552004"

Revision as of 05:07, 26 October 2020

hofM-hofN-hofO-hofP-hofQ(gene from DH5a)

HofM is a DNA utilization protein which is required for the use of extracellular DNA as a nutrient. Its functions include the carbon utilization and DNA catabolic process. HofN is a DNA utilization protein which is required for the use of extracellular DNA as a nutrient. Its functions include the carbon utilization, DNA catabolic process, type IV pilus biogenesis and type IV pilus-dependent motility. HofO is a DNA utilization protein which is required for the use of extracellular DNA as a nutrient. Its functions include the carbon utilization and DNA catabolic process. HofP is a DNA utilization protein which is required for the use of extracellular DNA as a nutrient. Its functions include the carbon utilization and DNA catabolic process. HofQ is a DNA utilization protein which is required for the use of extracellular DNA as a nutrient. Could be the porin responsible for transport of DNA across the outer membrane. Its functions include the carbon utilization, DNA binding, protein secretion and DNA catabolic process.This part is in the part collection where we have 13 genes that code for the proteins.

The part collection includes: Parts that are different kinds of type 4 pilus: BBa_K3552000 BBa_K3552001 BBa_K3552002. Parts that are the generator of the type 4 pilus: BBa_K3552003 BBa_K3552004 BBa_K3552005 BBa_K3552006 BBa_K3552007 BBa_K3552008 BBa_K3552018 BBa_K3552019 BBa_K3552020 BBa_K3552021 BBa_K3552022 BBa_K3552023 BBa_K3552024 BBa_K3552025 BBa_K3552026 BBa_K3552027 BBa_K3552028 BBa_K3552029. Parts that are a complete circuit: BBa_K3552009 BBa_K3552010 BBa_K3552011 BBa_K3552012.

Our part collection can instruct other teams to designed new rechargeable pilus and substitution of different major pilin.

Sequence and Features

Reference

Luna Rico, Areli et al. “Functional reconstitution of the type IVa pilus assembly system from enterohaemorrhagic Escherichia coli.” Molecular microbiology vol. 111,3 (2019): 732-749. doi:10.1111/mmi.14188

Usage and Biology

HofM is a component of the IM complex that is connected to the outer membrane secretin channel formed by the hofQ and allows pilus exposure on the bacterial surface. HofN is a component in the transmembrane protein that connected to the outer membrane secretin channel formed by the hofQ. HofO is a component in the transmembrane protein that connected to the outer membrane secretin channel formed by the hofQ. The inner membrane anchored assembly platform complex is connected via hofP to the outer membrane secretin channel formed by hofQ that allows pilus to expose on the bacterial surface. The hofMNOPQ operon encodes the assembly platform complex connecting HofB ATPase with the secretin hofQ. The hofMNOPQ is the generation pathway of pilA from the inner membrane to the surface of outer membrane.

Characterization

We first uses the bacteria DH5a as the template and obtain the DNA fragment of 12 genes. We verified the practicability of the generator through the western blot of pilA in vivo which confirmed the usage of the generator because the PilA is able to display outside the cell.