Latest revision as of 03:23, 25 September 2024

Tub2 promoter for Chlamydomonas reinhardtii (Phytobrick)

This basic part contains a combination of the Tub2 promoter and a 5'UTR (A1-B2) for Chlamydomonas reinhardtii and was built as a part of the Kaiser Collection. Combined with a CDS and a terminator, this level 0 construct leads to the expression of a target protein.

The Tubulin2 promotor was used as a promotor for the hygromycin resistance in the L2AB (BBa_K3002227) construct. The different promotor, compared to the one for the coding sequence, prevent repetition in the DNA which can decrease the expression of the constructs. The ligation of the resistance cassette was successful as well as the secretion of the substrate.

The Kaiser Collection

We are proud to present our very own MoClo part collection for C. reinhardtii - the Kaiser collection.

These 20 Parts are specifically designed and codon optimized for Chlamydomonas. Among them are regulatory elements, antibiotic resistances, resistance cassettes, secretion signals and tags. These parts were tested and optimized thoroughly and we can guarantee that they work 100%. With these, expression and secretion in Chlamy will be a success. Because this is a MoClo collection, the parts are highly standardized for worldwide application. The combination with other part collections works fast and easy. While in MoClo, nomenclature is a bit different from the iGEM BioBricks, it is quickly explained:

Level 0 parts are equivalent to basic parts, e.g. Promoters, coding sequences, etc.

Level 1 parts are combinations of basic parts and usually form functional transcription units.

Level 2 parts are combinations of Level 1 parts, in case you want to transfer multiple transcription units at once. For example, you can pair your gene of interest with a selection marker.

The great thing about the Kaiser Collection and MoClo is that the ligation works in a one pot, one step reaction, as the Type IIs restriction enzymes cut out their own recognition sites. This way, multiple constructs can be combined linearly in a fixed order to create complex structures. This is ensured by the standardized overlaps that assign the parts one of 10 positions in the final constructs. After trying MoClo once, you won’t go back to traditional ligation. It is incredibly easy and reliable. For this reason, we believe that our Kaiser Collection will strike a significant chord, as the future lies in standardized, easy to use methods such as MoClo. Visit our part collection site to get an overview over all parts of the Kaiser Collection

Sequence and Features

Usage and Biology

Team UVU-Utah-2, 2024

We obtained this part in the Chlamydomonas MoClo kit as described in Crozet et al. (2018). We utilized and characterized this part in composite part(s) K5078007. Please check out the documentation for that composite part for more details!