How Does Quorum Sensing Affect Bacteria

This pBlu lab had for purpose to present the changes of the strain of E. coli bacteria due to new genetic information being introduced into the cell. In this experiment we are freezing and heat shocking the E. Coli bacteria that is then forced to take the plasmid DNA. The E. coli then transforms the pBLu plasmid, which carries the genes coding for two identifiable phenotypes. After following the Carolina Biological steps our lab worked well and we able to see some colonies of bacteria on the plates. The x-gal plate showed a significant amount of bacteria to confirm that the pBlu plasmid took over the E. coli strain.

The pGLO plasmid is engineered to express green fluorescent protein (GFP) in the presence of the sugar arabinose as well as the ampicillin resistance gene β-lactamase (bla) (Brown, 2011). Original E. coli HB101 do not have ampicillin resistance or the GFP gene allowing them to glow under UV light. In this experiment, we transformed E. coli HB101 with the pGLO plasmid by heat shock to make the bacterial cells competent, allowing the plasmid to enter the cell (Brown, 2011). The mixture of bacteria with pGLO plasmid were given recovery time after heat shock, then spread on LB/amp and LB/amp/ara agar plates. The bacteria mixture with no plasmid added were spread on LB and LB/amp agar plates and all four plates were incubated at 37°C for

This experiment was designed to test and observe the transformation efficacy of the pUC18 and lux plasmids in making E. coli resistant to ampicillin. Both plasmids code for ampicillin resistance, however, the lux plasmid codes for a bioluminescence gene that is expressed if properly introduced into the bacteria’s genome. The E. coli cultures were mixed with a calcium chloride solution and then heat shocked, allowing the plasmids to enter the bacteria and assimilate into the bacterial DNA. The plasmids and the bacteria were then mixed in different test tubes and then evenly spread onto petri dishes using a bacterial spreader, heating the spreader between each sample to make sure there is no cross contamination. Each of the dishes was labeled and then incubated for a period of 24 hours. The results were rather odd because every single one of the samples grew. Several errors could have occurred here, cross contamination or possibly an error in preparation as every single sample in the class grew, meaning all samples of the bacteria transformed and became ampicillin resistant.

The experimental part of the lab consists of setting up the materials needed. A sample of E.coli and a solution of calcium chloride are first obtained and placed in different test tubes. 630µL of Calcium Chloride (CaCl2) are then removed from the test tube and inserted into the test tube containing E.coli cells (Alberte et al., 2012). The newly formed substance of Calcium Chloride and E.coli is then mixed and incubated in ice for 10 minutes, making the cells more competent. Two test tubes are obtained and labeled; the first test tube is labeled with pUC18 and the second one with “Lux” to represent the plasmids being used. These two test tubes are then incubated in ice. 3µl of the set plasmid are added to each of the two test tubes. The test tubes are tapped to guarantee the cells are well

In the pGLO Bacterial Transformation lab, Escherichia coli is transformed with a gene encoding green fluorescent protein by inserting a plasmid containing the GFP gene, beta-lactamase, and arabinose into the bacterium. Successfully transformed bacteria will grow in the presence of ampicillin and glow a bright green color under ultraviolet light. The sugar arabinose is responsible for switching on the GFP gene in the transformed cells, without it, the gene will not be expressed.

How does the addition of pGLO plasmid to a solution containing E. coli bacteria affect the growth and characteristics of the bacteria? Genetic transformation is the incorporation of foreign DNA into an organism to potentially change the organism’s trait. Plasmids are small circular DNA that replicate separately from the bacterial chromosome. In nature, these plasmids can be transferred between bacteria allowing for the sharing of beneficial genes. Due to this characteristic, plasmids allow for genetic manipulation and can be moved between bacteria easily. The pGLO plasmid utilized in this experiment encodes the gene for Green Fluorescent Protein (GFP), which under the right conditions can produce a glow. The gene regulation system present in the pGLO plasmid requires

This is proven by the survival of the bacteria in the LB/amp +pGLO dish, as the same bacteria that lacked the modification were killed by the ampicillin. However, we were unsuccessful in genetically transforming bacteria to be fluorescent. This is because in the LB/amp/arbo +pGLO dish, we were unable to locate any bacteria.

If a gene that codes for Green Fluorescent Protein transforms bacteria and GFP glows when transformation occurs, then when two micro test tubes have 250 microliters of transformation solution and places in an ice bath, then 2-4 bacteria colonies are added to each tube with a sterile loop; then a plasmid (pGLO) is added to one of the tubes, incubated in ice for 10 minutes, then heat shocked for 50 seconds at 42 degrees Celsius, then back into 9ice for two minutes; then LB nutrient broth is added to both tubes (250 microliters) and set out at room temperature for 10 minutes. Then, 100 microliters of each solution in the tube are added to four

Conclusion: The results from the bacterial transformation lab did support my hypothesis. My hypothesis said that the petri dish labeled +LB/AMP/ARA would have green glow bacteria. This was proven true as this particular petri dish had 34 bacterial colonies that glowed neon green when shown under UV light. One primary error I made was that I did not spread the bacteria as much as I needed to. This resulted in the bacteria being centered in the middle. In the future, I would take more time to ensure that the bacteria was completely spread out. Another error I made was that I applied the DNA before adding the bacteria. To prevent this mistake, I would take more time to do the

First, 500μl of calcium chloride was added to a DNA tube. Calcium chloride makes it easier for cells to transform. Approximately 15 E. coli colonies were then transferred to the tube. 250μl of the DNA was transferred into another tube, where 10μl of pGFP was then added. The plasmid for green fluorescent protein is what will be transformed into the E. coli’s DNA so it will express the GFP. The DNA tubes were then incubated in hot and cold water .This process is called “heat shocking” and affects the permeability of cells, allowing for new DNA to move into cells more easily. Initially the tubes were put in ice for 10 minutes, they were then put in water that was 42℃ for 90 seconds, after that they were put on ice for another 2 minutes, 250μl

They coordinate certain behaviors based on the local density of the bacterial population. Many species of bacteria use Quorum Sensing to coordinate gene expression. Quorum Sensing is a bacterial cell to cell communication process that involves the production, detection, and response to extracellular signaling molecules called autoinducers. Autoinducers collect in the environment as the bacterial population density increases. Quorum Sensing is just a process bacteria use to talk to one another.

Plasmids are small DNA atoms that are found in the nucleus region of a bacterium. They provide an extra assistance to a bacterium that allows them to survive and reproduce in certain environments, such as making the bacteria resistant to specific antibiotics. In the experiment being conducted, plasmids pUC18 and lux will be tested to determine the different types of growth with Escherichia coli present. In order to do this, CaCl2 was transferred into a tube with E.coli and incubated for 10 minutes. Different measurements of micro liters of various substances are added to the test tubes and later incubated for 15 minutes. A separate test tube is collected, labeled and added its corresponding micro liters of the competent cells. Once this is

Ampicillin is a beta-lactam antibiotic which can be used to treat a large number of infections. For example, Escherichia coli (E. coli) bacteria is terminated by this specific antibiotic. Ampicillin interferes with the formation of bacterial cell walls and thus kills newly dividing cells that must form new cell walls. Plasmids contain genes that create antibiotic resistance to their host cell. The pGlo plasmid contains an Ampicillin resistance gene. Therefore, bacteria that take up the plasmid and transform become resistant to Ampicillin. To carry out this experiment, my colleagues and I took four petri dishes containing the bacterial host cells (E. coli) on each and the Ampicillin on two of those four. Through a tedious process, we added the pGlo plasmid to one petri dish of just the host cell and one petri dish of the host cell and the Ampicillin antibiotic. When examining the results of our experiment, we noticed that both petri dishes containing zero Ampicillin entirely submerged in a lawn of the Escherichia coli bacteria, the petri dish containing no plasmid but some Ampicillin displayed zero growth, and the petri dish containing the plasmid and Ampicillin showed individual colonies of the E. coli bacteria glowing under a UV light. The reason this occurred is because the petri dishes that showed a lawn of bacteria contained no Ampicillin so there was no antibiotic to kill the bacteria off. The petri

Although traditionally thought to be a multicellular characteristic, unicellular bacteria populations behave cooperatively by means of small, diffusible chemical signals. The inter and intraspecies bacterial pheromone communication creates behavior synchronization within a species. The timing of a host infestation, genetic modification for niche adaptation based on a new energy source, or the production of a secondary defense metabolite are all examples of cooperative behavior that depend on intercellular bacterial communication (Everts, 2006).

Bacterial transformation is the process of moving genes from a living thing to another with the help of a plasmid.The plasmid is able to help replicate the chromosomes by themselves; laboratories use these to aid in gene multiplication. Bacterial transformation is relevant in everyday lives due to the fact that almost all plasmids carry a bacterial origin of replication and an antibiotic resistance gene(“Addgene: Protocol - How to Do a Bacterial