RNA Therapeutic Lab Report

_Alexandra DeWitt_____________ __5/1/13____________ Author Date _10_________ Lab Section # Increasing Sucrose Concentrations Causes the Osmotic Concentrations of Disks of Potato Tubers to Decrease Abstract: Introduction: The biological membranes are composed of phospholipid bilayers, each phospholipid with hydrophilic heads and hydrophobic tails, and proteins. This arrangement of the proteins and lipids produces a selectively permeable membrane. Many kinds of molecules surround or are contained within

Michelle Trujillo 5702361 Michaela Salisbury BSC 1010L U60 Effects of pUC18 and lux Plasmids on Ampicillin Resistance of Escherichia coli Abstract 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.

Experiment 1: Coding In this experiment, you will model the effects of mutations on the genetic code. Some mutations cause no structural or functional change to proteins while others can have devastating affects on an organism.

(Biology Dept.). 0.1 ml of E.coli K or 0.1ml of E.coli B was added to the 10 fold dilution. Using soft agar technique, the growth media mixture with E.coli was plated and incubated.

Discussion What was expected that the strains of E. coli that did not have a resistance to ampicillin would not grow. The transformed strain also changed to a blue color when the X-gal was present in plate. The transformed cells also grew because they were free of the ampicillin because they possessed the amp gene that they used as an shield against the ampicillin antibiotic. The transformed cell who turned blue did so because the gene converted the sugar to a blue color but also contained the amp gene to ensure that they grew even when the ampicillin was present. The growth of the colonies on the plates

Introduction: 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

The purpose of this experiment is to make E.Coli competent so that it can be transformed in order to become immune to ampicillin, then we would be able to determine the transformation efficiency of the culture. We determine this by preparing 4 plates of E.coli, each labeled “LB-plasmid”, “LB+plasmid”, “LB?Amp-plasmid”, and “LB/Amp+plasmid”. This meant that either should have lacked plasmid and Ampicillin, with plasmid but lacked Ampicillin, without plasmid but with Ampicillin, or were with Ampicillin and plasmid, respectively. Then we made the bacterial cells competent by adding CaCl2 to 2 vials of the colony (one with plasmids), and incubating on ice, then heat shocking, and returning to ice. Luria Broth is then added and left to sit for 5-15

E.coli outbreaks have steadily grown over the last few decades. An expansion in big farming has led to E. coli not only being found in meat, but vegetation as well, due to waste runoff. This has increased our need for adequate antibiotics that can fight bacteria, like E. coli. The

Instead, they argued in favor of the recombination dependent mutation theory. E.coli with a Lac- mutation were found to revert back into Lac+ bacteria when grown in lactose and minimal media. However, the mechanism was still unclear. Some scientists argued the lac region still had some ability to catalyze lactose, and will be amplified until it can grow on lactose. On the other hand, scientist at Indiana University state revertants are created through DNA error. Through three experiments, scientist argued against the amplification dependent mutation theory. Transposons were inserted at various distances from the lac region, and transformation frequency did not decrease as the transposon and lac distance decreased. Amplification of the transposon and the lac region should have occurred at the same frequency because they are close together on the plasmid. Furthermore, tetA gene, another gene on the plasmid, was also selected in the experiment. Similarly, the reversion of this gene was not tied to the reversion of the Lac- no matter the distance. Lastly, the amplification theory states alleles must be cis to undergo amplification. However, scientists found that reversion still occurs when the lac region is trans. Thus, these scientists debunk the amplification dependent mutation theory in favor of the recombination dependent mutation

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).

Escherichia coli, or E. coli, is a common bacterium that can be found in diverse environments all over the planet, including the gastrointestinal tracts of animals and humans. Many of these strains of E. Coli are essential mechanisms in the digestive tract, while others are pathogens that can cause complications

The transformation of E. coli using plasmid DNA was a success. The positive control plate had a near lawn of blue colonies growing on the plate. This indicated that the E. coli cells took up the plasmid and became ampicillin resistant. The blue colonies formed because

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For millions of years’ bacteria have been surviving to their environments and been at war against other microorganism. This wars have lead them to creating an unstoppable arsenal of weapons they can use to survive. Unlike virus that need a host cell to survive, bacteria can survive anywhere since they can share their DNA with each other. This allows bacteria to survive in places like radioactive waste, zero oxygen environments, and even in absolute darkness. Unlike human beings who are born with a specific genetic code, bacteria have the capability of changing that code. Bacteria have three methods of evolving themselves in order to survive the harsh is of conditions. The three methods are called horizontal gene transfer. The first is Transformation, but some pathologists call it the “Funeral Grab”.

To deduce if UV Radiation induced a mutation in the DNA of Serratia Marcescens and prevented the production of the red pigment called Prodigiosin. Hypothesis: Maybe the disappearance of red color in Serratia Marcescens is caused by the UV light which induces mutations in its DNA and is known as