Bacterial Art and GFP Nick Slawson

Bacteria refresher Bacteria are prokaryotic microorganisms (Small) found in practically all areas. Are shaped either as cocci (circle), bacilli (Rod), or spiral and most are either gram negative or gram positive. Used in research and biotechnological projects. Brief review of bacteria. Students should already have an understanding of bacteria including some examples of bacteria and how they affect people.

Transformation; DNA swapping Bacteria, while being single celled organisms, are able to acquire new DNA and new traits. This process is called transformation which is the ability for a bacteria to uptake DNA from its surroundings. This allows new genes to flow between bacteria and thus new proteins and new abilities to emerge from different strains of bacteria. Transformation should be new to them. Explain that bacteria also can change and produce new proteins even though they are single celled organisms. Talk about how bacteria become resistant to antibiotics, maybe ask why they think that happens before introducing transformation which is the reason why it can. Use the picture to really cement what can happen with transformation.

Transformation Explain Griffiths experiment here. Griffith took two strands of pneumonia (a deadly strand and a non-lethal strand, label them R and S or so for the students) and took turns placing each strand into mice. Eventually, he placed dead R, or deadly, pneumonia strands along with live non-lethal strand into mice and found that the non-lethal strands took up the lethal DNA and became lethal themselves.

Why Transformation? Transformation has been an incredible tool in biology as it has allowed scientists to insert new DNA into bacterium and cause genetic changes. Scientists have done many novel things with this, but mostly, they use it to mass produce plasmids or test new plasmids for future use. This is a slide that allows us to inform the students that what they are learning is very pertinent in biology right now. They are learning a skill that is used often in biology for a variety of reasons. It helps to connect the learning to the real world.

Plasmids Plasmids are a circular pieces of DNA that bacteria have that are separate from their chromosomal DNA. Bacteria can transfer this to acquire new DNA to make new proteins. We as scientists create plasmids to move new genes into existing bacteria. Plasmids are awesome! They are found in all three domains of life and are extremely helpful pieces of DNA. It is important to note that they are in factg SEPARATE from chromosomal DNA but they can replicate on their own also. We create plasmids for tons of things including making bacteria produce insulin and even glow as we are about to do!

Green Fluorescent Protein (GFP) Green fluorescent protein is a protein that is found in species of jellyfish (first isolated from Aequorea victoria) that causes a bioluminescence (glowing). This protein, as its name states, allow the jellyfish to glow when calcium is present in the water. GFP has since been used as a marker of expression in molecular biology. Make sure to note that these jellyfish glow when Calcium is in the water. We did extract the GFP gene from jellyfish, let them know that we can glow other things with the same gene. Also introduce pGlo which is the plasmid we made for the GFP gene.

pGlo, and the age of Fluorescent Proteins Aside from GFP, there are now many fluorescent proteins to choose from in creating markers for experiments. Along with that, a novel plasmid was created called pGlo which is used in biotechnology as a stepping stone for genetically modified organisms. Talk about pGlo as a plasmid and how it can be transformed into cells which causes green fluorescents under UV light. Talk about how this was huge for biology as a stepping stone for genetic modification and for use in molecular biology as a marker. This was Nobel Prize work! Students should also like the GFP cat photo.

Now its your turn! Today you will begin your own bacterial transformation. You will be able to pick a fluorescent protein and express it in E. coli causing it to glow the specified color. This will give you a glimpse into one of the most widely used techniques in science! It will also help you hone your sterile technique and lab skills.

Lets glow!