Difference between revisions of "Part:BBa K2918000"

Revision as of 07:13, 4 October 2019

P_BHR

DNA polymerase of the Φ29 bacteriophage

Overview

The replication of DNA and its conversion into functional proteins are vital processes in all living systems. DNA is copied during the replication process. The bacteriophage Φ29 contains a DNA replication machinery which replicates the linear plasmid by itself. This process is called orthogonal replication and can be beneficially used. The desired gene can be expressed in other hosts without interfering with the genome of its host. Our Sci-Phi 29 tool is based on the Φ29 DNA replication system and its four proteins. The terminal proteins (TP) cap the linear DNA, protect the linear DNA and are the starting point for the Φ29 DNA polymerase (DNAP/p2). DNAP binds to the TP and replicates the DNA. During the replication, single and double stranded DNA is protected by respectively single stranded DNA binding proteins (SSB/p5) and double stranded DNA binding proteins (DSB/p6).

Strain Construction

Aim: To clone the promoter in a level 0 MoClo backbone (pICH41233).
Procedure: The DNA sequence of the part was synthesized by IDT with flanking Bpi1 sites and respective MoClo compatible promoter overhangs. The cloning protocol can be found in the MoClo section below.

Sequence and Features