Bacterial Transformation using pGLO involving X and Y genes
Genetic transformation is due to a direct cause in the change by genes, due to the cell in taking and expressing traits from a separate piece of DNA. Naturally proficient bacteria are able to absorb exogenous DNA and go through genetic transformation. (Chen & Dubnau, 2004) The purpose of this experiment was to discover how a gene could be moved from one organism to a different organism with the help of plasmid. The cells that are capable of acquiring these traits from the other organism are known as being competent. (Weedman, 2013). In this particular experiment we will genetically transform the bacteria E.coli by inserting a gene through heat shock, this gene codes for Green Fluorescent Protein, also known at GFP. The GFP gene originally comes from a jellyfish and under an ultraviolet light the bacteria that acquired the gene with glow a brilliant fluorescent green color. (Portman et al. 2013). If the cells’ nutrient medium has the sugar arabinose added to it then GFP can be turned on. (Weedman, 2013). To determine if our hypothesis was correct, we used four differently prepared plates. The four plates each contained a different combination of the following; arabinose, ampicillin, LB nutrient broth, and pGLO plasmid. The combinations were; +pGLO LB/amp, +pGLO LB/amp/ara, -pGLO LB/amp, and –pGLO LB. Our hypothesis was: the plates with pGLO will have growth because they are resistant to the antibiotics involved, the plate with ampicillin and without pGLO will show no growth due to the fact that the antibiotic compromises the bacteria, and the plates that will grow will be the ones containing pGLO since they obtain the trait for glowing. Materials and Methods:
All methods were obtained from (Weedman, 2013)
Before beginning the experiment obtain latex gloves, two microcentrifuge tubes, a beaker filled with ice, a micropipetter, micropipetter tips, transformation solution...
Citations: Chen, Inês, and David Dubnau. "DNA Uptake during Bacterial Transformation." Nature.com. Nature Publishing Group, Mar. 2004. Web. 02 Dec. 2013.
Gregory, Michael J. "Bacterial Transformation Lab." Bacterial Transformation. N.p., Feb. 2004. Web. 02 Dec. 2013.
Portman, Drew, Sarah Micallef, and Matt Powell. "Genetic Transformation Using Plasmid Infused DNA of E. Coli Creates Ampicillin Resistance in Strain and Is Marked by the PGLO Luminescent." D. Portman Et Al. SCMED2031. /Molecular Cell Biology Practice, October 2013 (n.d.): n. pag. Print.
Weedman, Donna. "Chapter 10/Genetic Transformation." Life 102 Attributes Of Living Systems. E ed. Vol. 7. Minneapolis: Bluedoor, LLC, 2013. N. pag. Print. Lab Manual.
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