The Effects of pGLO Plasmid Injection on Prokaryotic Gene Expression.
By Kush Patel
3rd Period, AP Biology, February 12, 2024.
Lab Partners: What is the difference?
Brody Bonsor
Andrew Zhang
Marianne Baquero
Clarie Gabrielson
Abstract: In the pGLO lab, E. coli bacteria were genetically modified using a plasmid that contained three key genes: araC, GFP, and ampicillin resistance. Through a process of heat shock and incubation, the pGLO plasmid was successfully introduced into the bacteria. This allowed the transformed bacteria to express the GFP gene and fluoresce green under UV light, while also providing resistance to ampicillin. The success of the transformation was determined by comparing the number of colonies that …show more content…
The first indicator was bacterial growth, which was observed on four agar plates containing different combinations of LB broth, ampicillin, and arabinose. The second indicator was fluorescence, which was observed under UV light. The presence of fluorescence indicated that the bacteria had taken up and expressed the pGLO plasmid, which contained the GFP gene that produces green fluorescent protein.
The four plates used in the experiment were the +pGLO LB/AMP/ARA plate, the -pGLO LB/AMP plate, the +pGLO LB/AMP plate, and the -pGLO LB plate. The +pGLO LB/AMP/ARA plate contained LB broth, ampicillin, and arabinose, and was used to test for successful transformation. The -pGLO LB/AMP plate contained LB broth and ampicillin and was used as a negative control to test for contamination. The +pGLO LB plate contained LB broth and was used as a positive control to test for bacterial growth. The -pGLO LB plate contained no additives and was used as a negative control to test for bacterial …show more content…
coli) bacteria and saw successful bacterial growth under certain conditions. Only the +pGLO LB/AMP/ARA plate showed substantial bacterial growth and fluorescence under UV light. There was a difference in growth and fluorescence between the bacteria with the pGLO and the bacteria without the pGLO. This was due to the genes provided by the pGLO, which were the araC gene, GFP gene, and the ampicillin-resistant gene. The pGLO had to be uptaken by the E. coli bacteria to give it the GFP protein which allowed the bacteria to be fluorescent and also resistant to ampicillin due to the resistant gene in pGLO. Arabinose also needs to be present on the agar plate because arabinose activates a special promoter that regulates the expression of the GFP gene. Furthermore, these results support the alternative hypothesis that the pGLO plasmid will be transformed into the E.coli bacteria and be expressed. This was shown by how only the E.coli bacteria with the pGLO plasmid were able to survive and grow due to pAMP which gave the E.coli bacteria ampicillin resistance. This lab relates to course concepts like gene expression and bacterial transformation. Potential limitations to this experiment are not all bacterial cells may successfully take up the pGLO plasmid, resulting in a lower transformation efficiency than desired. This could have been caused by the difference in time in the
By Kush Patel
3rd Period, AP Biology, February 12, 2024.
Lab Partners: What is the difference?
Brody Bonsor
Andrew Zhang
Marianne Baquero
Clarie Gabrielson
Abstract: In the pGLO lab, E. coli bacteria were genetically modified using a plasmid that contained three key genes: araC, GFP, and ampicillin resistance. Through a process of heat shock and incubation, the pGLO plasmid was successfully introduced into the bacteria. This allowed the transformed bacteria to express the GFP gene and fluoresce green under UV light, while also providing resistance to ampicillin. The success of the transformation was determined by comparing the number of colonies that …show more content…
The first indicator was bacterial growth, which was observed on four agar plates containing different combinations of LB broth, ampicillin, and arabinose. The second indicator was fluorescence, which was observed under UV light. The presence of fluorescence indicated that the bacteria had taken up and expressed the pGLO plasmid, which contained the GFP gene that produces green fluorescent protein.
The four plates used in the experiment were the +pGLO LB/AMP/ARA plate, the -pGLO LB/AMP plate, the +pGLO LB/AMP plate, and the -pGLO LB plate. The +pGLO LB/AMP/ARA plate contained LB broth, ampicillin, and arabinose, and was used to test for successful transformation. The -pGLO LB/AMP plate contained LB broth and ampicillin and was used as a negative control to test for contamination. The +pGLO LB plate contained LB broth and was used as a positive control to test for bacterial growth. The -pGLO LB plate contained no additives and was used as a negative control to test for bacterial …show more content…
coli) bacteria and saw successful bacterial growth under certain conditions. Only the +pGLO LB/AMP/ARA plate showed substantial bacterial growth and fluorescence under UV light. There was a difference in growth and fluorescence between the bacteria with the pGLO and the bacteria without the pGLO. This was due to the genes provided by the pGLO, which were the araC gene, GFP gene, and the ampicillin-resistant gene. The pGLO had to be uptaken by the E. coli bacteria to give it the GFP protein which allowed the bacteria to be fluorescent and also resistant to ampicillin due to the resistant gene in pGLO. Arabinose also needs to be present on the agar plate because arabinose activates a special promoter that regulates the expression of the GFP gene. Furthermore, these results support the alternative hypothesis that the pGLO plasmid will be transformed into the E.coli bacteria and be expressed. This was shown by how only the E.coli bacteria with the pGLO plasmid were able to survive and grow due to pAMP which gave the E.coli bacteria ampicillin resistance. This lab relates to course concepts like gene expression and bacterial transformation. Potential limitations to this experiment are not all bacterial cells may successfully take up the pGLO plasmid, resulting in a lower transformation efficiency than desired. This could have been caused by the difference in time in the