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Team:Munich/Parts

Phactory

Parts Overview

Phactory Parts

Our parts collection consists of four basic parts and one improved composite part. All parts were generated during our project. Some parts relate to RecBCD enzyme & its inhibition and some to the translation detection via mTurquoise.

Parts Related to RecBCD

The RecBCD enzyme, also called Exonuclease V, is both a helicase that unwinds and separates linear DNA strands, and a nuclease that degrades linear DNA1. It consists of three subunits: RecB, RecC and RecD. RecBCD complex is essential for cell viability as it plays an important role in repair of DNA double-strand breaks by initiating homologous recombination2. However, RecBCD enzyme is also a highly processive at destruction of foreign DNA, like phage DNA3.

RecBCD is regulated by a cis-acting DNA sequence known as Chi (Crossover Hotspot Instigator) that activates its recombination-promoting functions. Interaction with Chi causes an attenuation of the RecBCD enzyme's vigorous nuclease activity4. Since Chi binds RecBCD with a greater affinity than other dsDNA substrate, this can be used to inhibit RecBCD activity in TX-TL, to more efficiently express linear DNA in TX-TL.

We provide basic parts containing the full coding region of the subunit RecC (BBa_K2722000) and RecD (BBa_K2722001), which can be used for an independent study of these subunits. Moreover, you can find the corresponding inhibitor Chi6 (BBa_K2722002) in our parts collection.

Parts Related to mTurquoise

The fluorescent reporter protein mTurquoise can be used for a variety of applications. For example, it can be used to measure translation efficiency. Our parts collection includes the coding sequence of mTurquoise (BBa_K2722003) for easy fusion to target proteins or cloning into other functional plasmids, as well as an expression construct of mTurquoise with a constitutive promoter (BBa_K2722004).

The parts are flanked by a BioBrick prefix and suffix and contain pSB1C3 as a backbone and come from risk group 1 organisms.

To learn more about our parts visit the Registry.

iGEM Munich BioBrick List

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