Endosymbiosis study kit by PCR of RuBisCO

Topics covered:
- Principle of PCR and study of endosymbiosis
- Study of the expression of two genes (nuclear and chloroplastic) encoding the enzyme RuBisCO
- Realization and analysis of DNA electrophoresis gel

Principles and objectives: Many scientific elements support the theory that chloroplasts are derived from a cyanobacterium integrated into an ancestral eukaryotic cell and that genes from the endosymbiotic bacterium have been largely transferred to the genome of the eukaryotic cell, constituting an example of horizontal transfer.
Can evidence be found of this transfer of genetic material from the chloroplastic genome to the nuclear genome of the plant cell? We propose to search for the presence of genes encoding the S and L subunits of RuBisCO in plants such as radish (Raphanus sativus) or cauliflower (Brassica oleracea variety botrytis) using the PCR amplification technique.
The experiment lasts 2 sessions of 60 minutes. The reagents are provided for 20 PCRs.

Composition: Ready-to-use primers, PCR mix [Nucleotides + Taq polymerase], DNA release, molecular weight marker, extraction buffer, PCR microtubes, micro-pins.
Additional equipment required: P20 micropipettes and tips, phorEasy electrophoresis system ref. 708150, Peltier module thermocycler ref. 591110, TAE buffer, agarose, GelGreen® dye

Comments: An experiment file is available on our digital platform to prepare the experiment, follow the protocol step by step, and investigate. The activity can be complemented by analyzing the nucleotide and protein sequences of RuBisCO in different species to partially reconstruct the evolutionary history of these species, or by researching the type of bacteria that associated with a eukaryotic cell and gave rise to the chloroplasts of plants and green algae. A ready-to-deposit DNA kit is also available on the same subject where students only need to analyze the electrophoresis, ref. 117231.