User talk:Pieguy314

FINAL PRODUCT FOR TRANSFORMATION LAB

1. Describe the results in the four agar plates and explain why these results occurred. The results of our experiment are as follows. The +pGLO/LB/amp agar plate contained a few white E. Coli colonies, but when placed under a UV light, they did not glow a fluorescent green. Because the agar plate did not provide the sugar arabinose with the nutrients the colonies did not glow. The colonies none the less grew even with the antibiotic ampicillin because a gene for ampacillin resistance was also located in the plasmid. The +pGLO/LB/amp plate produced many more white E. coli colonies and these colonies did in fact glow a fluorescent green color when placed under a UV light. This is because they included arabinose in their food supply which activated the pGLO gene. The -pGLO/amp agar plate on the other hand didn't show any growth because of the lack of the gene for resistance to ampicillin. The -pGLO/LB plate showed lots of growth of yellowish colonies, but again because of the absence of the pGLO gene, these colonies didn't glow when under a UV light.

2. What was transformed into the E. coli bacterial cells? Circular pieces of DNA called plasmids were transformed into the E. coli cells. These plasmids contained a gene for Green Fluorescent Protein (GFP) and a gene for resistance to ampicillin.

3. A selectable marker is a section of DNA placed on the DNA of interest to ensure that only bacteria containing this DNA live and grow. In our experiment what was the selectable marker and the DNA of interest? Our selectable marker was the ampicillin resistance gene during this experiment. The ampicillin in the agar plates were killed by the bacteria that hadn't taken up the plasmid. In this experiment the DNA of interest was the Green Fluorescent Protein or the GFP gene.

4. What is beta-lactamase? Beta-lactamase is a type of enzyme produced by some bacteria that is responsible for their resistance to beta-lactam antibiotics like penicillins, cephalosporins, cephamycins and carbapenems. These antibiotics have a common element in their molecular structure: a four-atom

5. What is DNA cloning? Did this occur in your experiment? When specific sequences of DNA have been isolated and placed into a recombant plasmid, bacteria is then allowed to replicate itself which produced million s of clones/ each clone now contains a dna sequence that was used for the protein research. It did occur in our experiment.

6. Look up and describe what an operon is. Specifically describe the arabinose operon and draw a diagram. An operon is a group of key nucleotide sequences including an operator, a common promoter, and one or more structural genes that are controlled as a unit to produce mRNA. Operons occur primarily in prokaryotes and nematodes. An arabinose operon is an operon containing arabinose, which is a monosaccharide containing five carbon atoms, and including an aldehyde (CHO) functional group. It has chemical formula C5H10O5. L-arabinose has the same configuration at its penultimate carbon atom as L-(-)-glyceraldehyde. Natural monosaccharides are nearly always dextrorotory and are chemically reducable to D-(+)-glyceraldehyde. With few exceptions, only in synthetic form would a levorotory L-(-)-arabinose be seen.