Antibiotic Resistance Pattern Of Parental Strains Essay

As predicted the E. coli colony transformed with either the PUC18 or the lux plasmid developed an ampicillin resistance. Which made it easier for them to not only survive but also replicate in both the LB agar plates and the LB ampicillin rich agar plate. However the E. coli colony not treated with the plasmids could not survive and colonize in the LB ampicillin rich agar plates. The plate that had no ampicillin in its environment and no plasmid treated E. coli served as a positive control for this experiment because it demonstrated how the E. coli would colonize and grow in a normal setting. The cells in the positive control plate grew into lawn colonies because they were not placed into a selective environment or transformed, so they had no need to acquire ampicillin resistance. Two plates in the experiment contained E. coli cells that were transformed with either the PUC18 or the lux plasmid but were placed in an ampicillin free environment. These two colonies grew

The Kirby-Bauer antibiotic tests indicated that the unknown is susceptible to chloramphenicol and ampicillin and resistant to penicillin G and tetracycline. The disinfectant sensitivity tests indicated that C. Freundii is resistant to all four of the disinfectants.

70µL of competent E.coli are added to both test tubes; pUC18 and Lux (Alberte et al., 2012). Both test tubes are then tapped and placed back into the ice bath for 15 minutes. While waiting, another test tube is obtained, filled with 35µL of competent cells and labeled NP for no plasmid. A water bath is preheated to 37 degrees Celsius and all three labeled test tubes are inserted into the bath for five minutes (Alberte et al., 2012). Using a sterile pipet 300µL of nutrient broth are inserted into both the control and Lux test tubes and 150µL are inserted to the no plasmid test tube to increase bacterial growth. All three test tubes are then incubated at 37 degrees for 45 minutes. Six agar plates are obtained and labeled to correspond each test tube, three of the plates contain ampicillin. A pipet is used to remove 130µl from each test tube containing a plasmid and insert it into the corresponding agar plate. For this, a cell spreader is first

The bacteria has ampicillin resistance gene and can either have no RNAi insert (control plate) or PCR product that is a portion of target gene (unc-22).

During the purification section of this lab, the LB/amp/ara agar plate was examined for well-isolated green colonies and the LB/amp plate was observed for white colonies with space between each other. These colonies were circled on the outside of the plates using a marker. Next, two 15 milliliter culture tubes containing 2 milliliters of nutrient growth media were obtained and labeled “+” and “-“. Using a new inoculation tube, the circled colonies from each plate were scooped out and immersed in their respective culture tubes. Once the bacteria was mixed into the solution, the tubes were sealed and placed horizontally into the 32⁰ incubator for 24 hours.

For this experiment, E. coli was best for genetic engineering because of their size, and their fast reproduction (Spilios, 2017). E. coli will be genetically transformed using an engineered plasmid. A plasmid is a circular piece of DNA which independently replicates and multiplies because it has its own origin of replication (Spilios, 2017). The pGLO is the plasmid used in this experiment. Plasmids are used as vectors and they contain manipulated genes such as genes coding for antibiotic resistance for drugs like ampicillin. This antibiotic resistance of such serves as the selectable marker in genetic transformation and for genetic transformation to proceed, the cell must reach competency which is the physiological state that is required for the vector plasmid to get into the cell for transformation (Spilios, 2017). While competency can be reached naturally in some organism, it must be reached artificially in E. coli through treatment with CaCl2 and exposing them to heat shock using incubation (Spilios, 2017).

Coli. The first standard E. Coli has no resistance plasmid while the second strain contains a resistance plasmid with genes protecting it from ampicillin. This standard E. Coli and pAMP (plasmid-Ampicillin) E. Coli were each streaked across plates containing the antibiotic and containing growth supportive Lurithea Broth. The purpose of this lab was to test their growth in each medium. Our hypothesis was that while the ampicillin resistant E. Coli would show growth in both LB and LB-AMP plate, the standard E. Coli would only grow in the LB plate for it contains no resistant plasmids against the

One person each in a team of four heavily inoculated two Mueller-Hinton agar plates with one of the cultures listed by aseptic transfer from a broth culture using a sterile cotton swab. Each plate was marked off into four segments, a total of eight sectors. One McFarland standard disk containing one of the eight antibiotics tested was placed, using

After analyzing the data recorded for both the agar plates containing ampicillin and those that did not, it can be concluded that the data provides enough evidence to reject the null hypothesis. There is enough evidence to support the alternative hypothesis stating that there is a correlation between plasmids coding for an antibiotic resistant gene and bacterial growth in ampicillin. When a bacterial solution containing either pUC18 or the lux plasmid is transformed in an agar plate containing ampicillin, only those cells which took in the plasmid are able to survive and replicate, forming individual colonies. Not all cells are transformed though, the chances of a successful transformation were extremely low. In order to see which cell transformed the cells were tagged according to their plasmid, in the plates containing pUC18 the only

In the BIOL2202 bacteria genetics practical, our results can hardly discriminate which sex mechanism was employed by E.coli to revert back to prototroph as the donor(JC158) and recipient strands(AB1157) were added into a Erlenmeyer flask and mixed together to facilitate conjugation. Besides, the reversion could also be attributed to transformation due to unknown transforming factors which somehow mutated the gene back to wild type configuration. When mixing began, both transformation and cell conjugation were possible, thus it was hard to differentiate which mechanism that ultimately leads to the appearance of prototroph.

Antibiotics are amongst the most important medical discoveries and their introduction represents a remarkable success story (Hedin, 2011). The term antibiotics literally means against life (Walsh, 2000). Thus antibiotics can be used against any microbe such as bacteria, viruses, fungi, and protozoa. However, some people use the term to only apply to bacteria, but in this paper, the more appropriate term will be used.

100 μl of 10-5 and 10-6 dilutions of donor cells were each plated onto MacConkey (MAC) agar plates without streptomycin. 100 μl of 10-5 dilution of donor cells and 10-5 and 10-6 recipient were also plated onto MAC plates with streptomycin. 100 μl of 10-4 and 10-5 dilutions of the conjugation mixture cells were plated onto MAC agar with streptomycin. All seven plates were inverted and placed in a 37˚C incubator for about 24 hours. The bacterial colonies on each plate were counted the next day (colony counts seen in Table I).

Bacillus subtilis is rod-shaped and has the ability to form a tough, protective endospore, allow the organism to tolerate extreme conditions. Some uses of Bacillus subtilis includes using as a model organism for laboratory studies. It can also be genetically modified to convert explosives into harmless compound of nitrogen, water and carbon dioxide. (Wikipedia, 2010)

In this laboratory experiment, we was introduce to an introduction to streaking and spreading of bacteria in agar plates such that single cells can be isolated from one another, each cell can reproduces to form a visible colony composed of genetically identical clones. Streaking and spreading bacteria is to obtain individual colonies is usually the first step in genetic manipulation of microorganisms.

This experiment was performed to test the hypothesis if LB nutrient broth, +pGLO and -pGLO Ampicillin, and Arabinose was placed in the E. coli plates, then there will be a significant growth in the newly transformed bacteria and it will possess the ability to glow under UV light. The measurements were recorded from the bent glass tube in each glass test tube. The transformation protocol tested for the newly possessed traits in E.coli bacteria. Throughout the experiment there were many probable reasons for failure. If the pipettes and sterile loop were not thrown out in between each use, a cross contamination could cause a miscalculation in the experiment causing the data results to fail. The hypothesis that was tested was validated due to the positive results with each experiment stating that newly transformed organisms due in fact pass on traits.