Hey people! Today BPT practical was great because it was a very short one! J We transferred several colonies of GFP(Green Fluorescent Protein) transformed E.coli from a fresh LB/Amp/Ara plate to the flask containing 100ml LB medium with ampicillin. This will be used to inoculate the fermentor for scale – up fermentation.
For a clearer understanding, please look at the video below.
I know what is running through your mind... "What is GFP" and "how it is incorporated in E.coli" rite? With Meena and Shu Fung here why worry?! We’ll tell you the answers!:P The green fluorescent protein (GFP) is bascially a protein comprising of 238 amino acids which originated from the jellyfish Aequorea victoria. The structural shape of the GFP is a beta barrel, a cylindrical anti-parallel beta sheet comprising of 6 alpha helices and 11 beta strands interconnected together by loops. By now... all of you should know what those complicated words mean…..and thanks to Mr. Michael for that! But if you are still blur... Here is a picture to make your life easy!
(Source: A website that most of us cope introduction for reports.... WIKIPEDIA!)
What’s so good about GFP that it is used in research is its ability to fluoresce green when exposed under UV light. Cool right?! Due to the usefulness of GFP, it is used as an indicator and as a bioreporter by the scientists.
Now... to the part where how GFP is incorporated in E.coli...
The GFP gene is incorporated in E.coli by first inserting GFP gene into the plasmid DNA known as the pGLO. The pGLO protein also contains antibiotic resistance gene in it which will only allow transformed bacteria to grow in an agar containing antibiotic like ampicillin. Subsequently, transformation solution is used to heat shock the pGLO plasmid into the E.coli cell. Heat shock is often done to increase the permeability of the cell membrane and allow uptake of the pGLO plasmid into the cell. Following that, the cells are allowed to grow such that it will eventually express the GFP gene and form protein. (It’s sooooooo MBT!)
We are not finished yet!! We found a cool picture on GFP on the net….and we want to share it with you!... Imagine if a GFP gene is engineered…. and there are mutants obtained from it... When u streak it on a plate... How do you think the outcome will look like? Can you picture it???
This is how it looks like!!! Bet you didn’t imagine this even in your wildest imagination!!:P It is a San Diego beach scene drawn with an eight color palette of bacterial colonies expressing fluorescent proteins derived from GFP.
