Revolutionary studies in the new fields of genetically modifying organisms have led humans to believe the world's starvation, low food production and other problems can be solved. GMO's are made through the process of taking a tumor inducing (TI) plasmid from a bacteria and inserting the gene of interest into it. The plasmid then undergoes a transformation as the agricultural bacteria grows and replicates its DNA. The bacteria is put into a plant cell, which then is obviously developed into what we call a GM plant. To identify these GMO's, it takes supreme skill, which is why I am doing this because I am a Biotech God. In the first day of our lab we will first be extracting the DNA from the plant cell. A 99 degree C water bath is used to break the cell wall, the plasma membrane and the nuclear membrane. Then, the added instagene matrix beads will kill the lurking DNAse that is ready to harm the newly exposed DNA, which becomes a wounded target. DNAse, as a side-note, is within the cell's cytoplasm because it protects the cell from foreign invaders who try and expose their DNA to the cell's. In the 2nd day, we will be attempting to amplify (produce in mass quantities) a piece of this DNA for testing. To do this, we will be using PCR, or a polymerase chain reaction. In PCR, what happens is you take a piece of our DNA, (target DNA), then identify the nucleotides within it. A primer, which is complimentary to the start of the sequence of the target DNA, usually in this case, a GM primer, is added to find this gene of interest. This primer can be though of as a sort of "heat seeking missile" that helps us find our GOI. On the Third day, we will be running a gel in gel electrophoresis. In lane 2, we have our GM primer, and in lane 1, we have our plant primer. These controls solve the issue, if one, of an error occured after viewing the gel, because there is no way of telling if the lab just didn't work, or if it's just not a GM food. If lane 1 shows up, we know gel electrophoresis didn't fail. If lane 2, when lane1 is banded, is there or not, determines if the food is GM or not.
Based on the results above, though blurry, one can notice that both bands showed up for every pair of lanes. However, lane 3 did not show up, therefor, in our case, our carrot that we tested was not GMO. During the course of this lab, I learned that humans can begin to test DNA through a PCR complex to discover the differences between DNA's and how maybe we can learn to test for more important things like hereditary diseases, or chance of having a disease. Genetics is our future, and we need to learn as much as possible to understand it, yet keep it under a contained limit.It cold benefit society positively, in so many ways.
Genetic Transformation will be the primary objective of this lab, which involves the insertion of a gene into another to alter its genetic make-up, usually for a particular phenotype change. Since genes are universal in all organisms, they can be carefully tampered with to transform a particular organism's trait. For example, in agriculture, if you take the gene that codes for drought resistance (maybe from a cactus) and insert it into a corn cell's DNA, then the corn cell will in effect produce baby corn with the drought resistance gene. In the world today, this can be very controversial. This topic goes into depth for a number of reasons but one negative effect of this might include the limited genetic diversity of the certain species.
Specific to this lab, we will be using the gene from the sea jelly (Aequorea Victoria) that encodes for the Green Fluorescent Protein, or GFP. After the insertion of the gene into the bacterial plasmid, or circular piece of DNA, the bacteria should hopefully glow a fluorescent green under ultraviolet light, like the sea jellies do. Plasmids in bacteria are essential to their survival. Usually, plasmids contain one or more genes that code for certain traits that can be beneficial to their development. Bacteria in nature can naturally transfer plasmids from cell to cell, which also explains the recent occurrence of antibiotic resistance. Bio-Rad's pGLO plasmid which we will be using has the GFP coding gene, as well as an ampicillin resistance gene. However, when inserted into the bacterial DNA mix, the pGLO doesn't just light up the bacteria. The addition of a control, a sugar aribinose, must be added to "switch on" the GFP gene. Without the sugar, the brilliant glow will not show.
Some possible sources of error include interferences from the environment such as spores or fungi for example, or if the setting is right for the proper bacterial growth.
The results were extraordinary, yet somewhat depressing as well. As usual, Mr. Chugh's sample is perfect, yet the rest of the classes' bacteria was stunted in growth by the very low temperatures over the weekend. The environment was not ideal for the bacteria to grow, so most of our specimen were destroyed. However, under nothing but ultraviolet light, Mr. Chugh's bacteria were glowing! They were shining brighter than I could've imagined.
