1. Are Euglena plants or animals?

2. NEITHER! Euglena are protists: a single celled, eukaryotic organism Some euglena have chloroplasts Some euglena are highly mobile heterotrophs So…..let’s revise the question…

3. Are euglena more closely related to plants or animals?

4. We can answer this question using the amino acid sequence of a particular protein We must pick a protein that all organisms being studied have in common (we will be using cytochrome c)

5. I. Retrieving amino acid sequences Organism name and protein name go here!

6. Search results: There are 12 matching proteins, click on this button to see the results

7. Protein results Select the protein that most closely matches the one given in your directions!

8. Getting the amino acid sequence: converting it to FASTA format

10. We will compare 2 species of Euglena to 2 species of plant (rice, Arabidopsis thaliana) and 2 species of animal (monkey, mosquito) I found the amino acid sequences from Entrez in the same manner and pasted them all into the same wordpad file

11. Edit the sequences; clear the first line (leave the >), and enter the species’ common name >Euglena Viridis GDAERGKKLFESRAGQCHSSQKGVNSTGPALYGVYGRTSGTVPGYAYSNANKNAAIVWEDESLNKFLENP KKYVPGTKMAFAGIKAKKDRLDIIAYMKTLKD >Euglena gracilis GDAERGKKLFESRAAQCHSAQKGVNSTGPSLWGVYGRTSGSVPGYAYSNANKNAAIVWEEETLHKFLENP KKYVPGTKMAFAGIKAKKDRQDIIAYMKTLKD >Arabidopsis MASFDEAPPGNAKAGEKIFRTKCAQCHTVEAGAGHKQGPNLNGLFGRQSGTTAGYSYSAANKNKAVEWEE KALYDYLLNPKKYIPGTKMVFPGLKKPQDRADLIAYLKESTAPK >Rice MASFSEAPPGNPKAGEKIFKTKCAQCHTVDKGAGHKQGPNLNGLFGRQSGTTPGYSYSTANKNMAVIWEE NTLYDYLLNPKKYIPGTKMVFPGLKKPQERADLISYLKEATS >Silvered leaf monkey MGDVEKGKKILIMKCSQCHTVEKGGKHKTGPNHHGLFGRKTGQAPGYSYTAANKNKGITWGEDTLMEYLE NPKKYIPGTKMIFVGIKKKEERADLIAYLKKATNE >Mosquito MGVPAGDVEKGKKLFVQRCAQCHTVEAGGKHKVGPNLHGLFGRKTGQAAGFSYTDANKAK GITWNEDTLFEYLENPKKYIPGTKMVFAGLKKPQERGDLIAYLKSATK

12. II. Creating the tree Copy and paste your sequences from notepad into ClustalW Click “execute multiple align”

14. Results page Scroll to the bottom Select n-j tree and click execute