The main objective of genetic transformation is to alter an organism’s traits. However, before any alteration can be made in an organism or any change can be detected, an all the way through examination must be made. In the colonies of E. coli on the starter plates, there are observable traits or characteristics such as color and number of colonies, distribution of colonies.
When the UV light has shone into the sample of the original pGLO plasmid DNA, the plasmid sample did not fluoresce.
On the LB plates, some E. coli are growing which do not contain ampicillin. The bacteria that did not receive the plasmid are growing on a plain LB plate.
With regard to the bacterial growth on each plate, there are numerous colonies on both LB/amp and LB/amp/ara plates. However, there’s a lawn of bacteria on the LB (-) pGLO plate.
There’s no growth on the LB/amp (-) pGLO plate.
As to the color of the bacteria, it is whitish on the (+) pGLO/amp plate and the (-) pGLO plates. However, the bacteria on the (+) pGLO/LB/amp/ara plate become visible to be white in color when exposed to normal, room lighting, but fluoresce brought green when bring to the long-wave UV light.
The spots being observed on each plate are optionally 75 colonies on the two (+) pGLO plates. The lawn of bacteria present on the LB plate contains a uniform dispersion of bacteria and individual colonies can not be counted.
It was observed that non-transformed traits include color of the bacteria which are whitish in color and colony size which are similar on both before and after transformation. In addition, of the E. coli traits the color and the ampicillin seemed to be significantly different after performing the transformation procedure. With regard to color, the colonies on the LB/amp/ara plate fluoresce green under UV light and the transformed colonies can grow on ampicillin resistance.
In order to determine how E. coli are affected by ampicillin through the use of two LB nutrient agar plates, some E. coli, and some ampicillin, same quantity of E. coli cells could be plated on two different LB nutrient agar plates containing LB nutrient agar and LB nutrient agar ampicillin, separately. There are observation and implications regarding the growth of the E. coli comparatively. If ampicillin has a negative effect on the growth of E. coli, then there should be fewer bacteria on that plate. If ampicillin has no effect, equal numbers of colonies on both plates would be observed.
With regard to the indication about the effect of ampicillin on the E. coli in the experimental result, it is expected that there should be few, if any, bacterial present on the ampicillin plate for antibiotics usually kill bacteria restrain their growth. The existence of any colonies on the ampicillin plate implies that those bacteria are tough to the antibiotic ampicillin.
In the two plates, bacteria that look like the non-transformed E. coli is expected to be found on the LB /(-) pGLO plate. Since these bacteria were removed form the starter plate, they would not have any plasmid added to them, and were replated on the LB plate. Hence, they were very similar to the non-transformed starter E. coli.
The transformed cells are found on the LB/amp/ara plates. Genetically transformed cells have taken up the pGLO plasmid and state the ampicillin resistance gene. The gene could live on the plate containing ampicillin. Thus, it is expected that there would be genetically transformed bacterial cells on the plates which contain ampicillin.
In order to determine if any genetic transformation has occurred, the LB/amp (-) pGLO and the LB/amp (+) plates should be considered for direct comparison. Cells that are not expressing the ampicillin resistance just implies that these cells were not treated with DNA (-pGLO) and would not grow on the LB/amp plates. Cells which were treated with DNA (+pGLO) should have the pGLO plasmid and should express ampicillin resistance gene. The corresponding LB/amp plate will have the transformed bacterial colonies.
If the genetically transformed cells get the ability to survive in the presence of the antibiotic ampicillin, other genes on the plasmid that have to do with transformation procedure, the plasmid must express a gene for ampicillin resistance.
The laboratory experiment is known as the genetic transformation. Genetic transformation refers to the when a dell takes up and express a new piece of genetic material – the DNA. This kind of transformation provides the organism with new traits identifiable after the transformation. With the pGLO transformation kit, students use the simple procedure in transforming bacteria with a gene that codes for Green Fluorescent Protein (GFP).
This activity will help students learn about the process of moving genes from one organism to another. This pGLO transformation kit provides the opportunity for an additional activity involving purification of the recombinant fluorescent protein from transformed bacteria using the GFP chromatography kit. The major purpose of this lab is to show among students visible changes in E. coli that have been transformed with a gene. Win this experiment, we have would know that the use of green fluorescent enables fast determination of protein expression
Komives, C., Rech, S., & Mcneil, M. (2004). Laboratory Experiment on GENE SUBCLONING For Chemical Engineering Students. Retrieved 28 December, 2007, from http://www.engr.sjsu.edu/ckomives/Courses/Biochemical%20Engineering%20Laboratory/additional%20materials/published%20article.pdf
Reynolds, J. (2004). Bacterial Transformation. Retrieved 28 December, 2007, from http://www.rlc.dcccd.edu/mathsci/reynolds/micro/lab_manual/transformation.html