Team:Lethbridge/Parts

The Next vivo system is made up of >40 unique parts encoding both protein and RNA components. The main focus of our team this year was to submit all 38 TX-TL protein components to the registry as a collection of parts that anyone could use to produce their own Next vivo system. Several of these TX-TL components were already in the registry but were under different expression control, lacked purification tags, and/or were not codon optimized for expression in E. coli. As such we sought to improve these parts for future use.

In addition to the TX-TL proteins, we also sought to BioBrick the ribosomal RNA genes and twenty tRNA genes. As a proof of concept for our novel tRNA purification design we only synthesized one representative tRNA gene before ordering them all. Additionally, the Next vivo system makes use of a previously submitted MS2BP part (BBa_K2109108).

Part Collection

The Next vivo part collection consists of the 38 TX-TL proteins required for RNA transcription and protein synthesis. Each part in the collection has a hexahistidine tag for simultaneous purification of all components, the T7 promoter (BBa_I719005), medium RBS (BBa_B0034), and double terminator (BBa_B0015).

Parts Submitted

This season we were able to submit 9 parts in pSB1C3 to the registry from our part collection and aim t submit more as we successfully ligate them into pSB1C3.

Composite Parts

General Protein Construct

General RNA Construct

This year we designed 41 composite parts. Of the 41 parts, it contains 38 proteins essential for prokaryotic transcription and translation (TX-TL) as well 3 additional constructs for tRNA purification and TX-TL validation.

Improved Parts

Of our 38 TX-TL components, five were already in the registry of standard biological parts. We decided to improve these parts for future use in our Next vivo system and for general use by others. Our improvements are documented here.

Basic Parts

All basic parts pages created on the registry of standard biological parts. Contains all 38 proteins essential for prokaryotic transcription and translation (TX-TL) as well as the parts for our tRNA purification and TX-TL validation construct.

Previous Parts Used

During the design of our project we utilized several parts already in the registry in our composite parts including: