Salmonella Bacteria For Producing Protein Of Interest And Rate Growth And Protein Concentration After Inoculate The E.coli Culture

(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.

When lactose becomes available the genes encoding β-galactosidase and lactose permease are upregulated in E. coli.

Every year over 96,000 people contract E.Coli, 3,200 of those people are hospitalized for E.Coli, and over 31 people die each year from E.Coli (Food News). E.Coli (otherwise known as Escherichia coli O157:H7) is a disease that is spread in many ways, the most common way being raw and uncooked food in the restaurant being served to the customer. Another way E.Coli is spread is through contact human/animal feces. E.Coli can be prevented, here are some ways to do so; Cook all meats to at least 160 degrees fahrenheit, wash your hands with warm water and soap (especially after touching raw meats), wash off all kitchen supplies thoroughly, use only pasteurized dairy and juice products, use/drink treated water, If you travel to another country that may have unsafe water don’t use tap or ice water, avoid raw fruits and veggies (Web MD).

Introduction: Transformation is used to introduce a gene coding for a foreign protein into bacteria. Hydrophobic Interaction Chromatography (HIC) is used to purify the foreign protein. Protein gel electrophoresis is used to check and analyze the pure protein. Research scientists use Green Fluorescent Protein (GFP) as a master or tag to learn about the biology of individual cells and multicultural organisms. This lab introduces a rapid method to purify recombinant GFP using HIC. Once the protein is purified, it may be analyzed using polysaccharide gel electrophoresis (PAGE).

Many people think of Escherichia coli as only a severe intestinal illness caused by eating uncooked, contaminated water, or unwashed fruits, but it is much more than only a negative (Amenu et al., 2014). The quick regeneration time of Escherichia coli makes it extremely useful in laboratory studies. In terms of research, a useful aspect of Escherichia coli is the extensive amount of information we have on them (Archer et al., 2011). Scientists have used Escherichia coli to host proteins, and it has been excellent resource for evolution studies as the bacteria has adapted frequently over the years (Archer et al., 2011). The non- pathogenic strains of Escherichia coli are frequently used in medical investigations involving diseases that are difficult to work with. Examples of these diseases include cancer or anthrax poisoning (Das et al., 2013).

An infectious disease that I would to address is salmonella, which is a waterborne pathogen disease or food poisoning. And the agent of the disease is a bacteria. It is commonly caused by eating contaminated food or drinking contaminated water and touching infected animals and not washing your hands afterwards (Centers for Disease Control and Prevention, 2015a). Salmonella infection especially affects the intestinal tract of a human body and live in animal and human intestines. The people primarily affected by the disease are zoologist, veterinarian or zoo keeper, old adults, infants, pregnant women and their unborn babies, older adults and a person with weakened immune systems. According to the Centers for Disease Control (CDC), Salmonella may be found in the feces of some animals, and people can become infected if they do not wash their hands after contact with animals or animal feces.

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a pathogen commonly associated with localized intestinal infection in humans. For its survival and propagation as a facultative intracellular pathogen, this Gram-negative bacterium has developed a variety of ways to manipulate host cell processes (Malik-Kale et al. 2011). Besides phagocytic internalization (by, e.g., macrophages, neutrophils, or dendritic cells), Salmonella infection is characterized by the bacterial-induced uptake of the pathogen into non-phagocytic enterocytes of intestinal epithelium (Perrett & Zhou 2013).

Each year, the Centers for Disease Control and Prevention (CDC) estimates that one in six Americans will become sick, hospitalized, or die as a result of foodborne illness (http://www.cdc.gov/foodborneburden/). With these kinds of statistics, food scientists are constantly developing new ways to prevent microbial contamination in foods, while regulatory agencies continue to educate the public on safe food handling practices. Food science is the study of the physical, chemical, and biological properties of food and the processes associated with them. An important aspect of food science is reducing or preventing the formation of harmful microorganisms in foods, through processes based on scientific research. Salmonella typhimurium (S. typhimurium),

E.coli O157:H7 bacteria was discovered in 1885 by a German scientist Theodor Escherich, his discovery also found that strands of the bacteria caused infants diarrhea and gastroenteritis which made this an important health discovery.” E. coli is one of the most frequent causes of many common bacterial infections, including cholecystitis bacteremia, cholangitis, urinary tract infection (UTI), and traveler's diarrhea, and other clinical infections such as neonatal meningitis and pneumonia.” Says the food poison journal, All You Need To Know About E. coli. The bacteria was first called bacterium coli but then was changed to Escherichia coli to honor the scientist that discovered it. There are over 700 serotypes of E. coli that are identified and not all of the ones identified are necessarily cause diseases in us humans, some help us and some give us infections. This disease is responsible for a big amount of contaminated foods and drinks. Foods that are

A future experiment for site directed mutagenesis/PCR might involve a slightly increased amount of template DNA and primers in the PCR reaction or increased amount of PCR product that is being transformed, in case if the transformation of the plasmid did not work. Another possible change could be done to the PCR thermal cycles, by altering the denaturation and annealing temperatures to a reasonable degree. This is to ensure amplification of the desired plasmid DNA incorporating the desired mutation with accuracy at the appropriate locations within the DNA template. A future experiment for protein expression and purification should be done with further precision. And more care needs to be taken as to include all necessary antibiotics during transformation and expression, depending on the type of plasmids used in the experiment. This is essential to obtain the desired protein containing the appropriate genes to procced with further

Bacteria belongs to a group of organism that lacks cell nucleus and membrane bound organells. This group of organisms are termed as prokaryotes. Prokaryotes follows the central dogma of molecular biology first proposed by Francis Crick in 1958 to synthesize proteins from mRNA through a process called translation and the mRNA is being synthesized from the DNA by another process called Transcription. Temperature, nutrient availibity are some key factors that start the process of synthesizing proteins in response to these key factors. Example. This paper will provide an explanation as to how bacteria decode the genetic information to produce proteins.

Salmonella enterica (formerly Salmonella choleraesuis) is a rod-shaped, facultatively anaerobic, flagellated, Gram-negative bacterium, a member of the genus Salmonella, belonging to the family Enterobacteriaceae. A number of its serovars are serious to human as well as animal pathogens. Among these sarovars, Salmonella enterica serovar Typhi is one of the most important food-borne pathogen causing typhoid. The genome of S. Typhi has a clonal nature but variations have yet been observed among its strains. Though a number of attempts have been made but success has yet not been achieved in controlling the disease caused by this bacterium. Salmonellosis is a condition that affects an estimated 2 million Americans each year, is common throughout

Next we turned regular E.coli cells, into chemically competent cells. This means that the cells were now able to act a host cel1 and accept the vector. Through transformation, we were then able to insert the pCS2 + GFP into the E.coli cells.. We then isolated the pCS2 + GFP from the chemically competent E.coli cells that we previously made, using a miniprep protocol. To observe whether or not we successfully isolated the pCS2 + GFP, we ran a gel. Figure 2 shows us that each sample contained the pCS2 +GFP after the miniprep was preformed. This supports that the pCS2 + GFP were all successfully

Salmonella is a gram-negative bacillus that causes inflammation of the GI tract and in some cases, if the immune response is not sufficiently powerful and treatment is not administered, can become systemic and cause even more serious conditions throughout the body. After ingestion, these bacteria cause infection by invading the epithelial cells of the small intestine and macrophages. Though there are more than two thousand different subspecies of Salmonella, few of them are able to cause serious conditions in humans—for most, the disease resolves itself in a matter of days. Those who are most affected by Salmonella infection are infants, the elderly, and people with compromised immune

Each well in ELISA plate was coated in triplicate with cell lysate and protein and incubated at 4°C overnight. Subsequently, blocking buffer containing PBS/ skim milk 3% (w/v) were used for 1h at 37° C and purified phages acquired from each round of panning were added to the wells and incubated at 37°C for 2h. After three consecutive washes with PBST and three washes with PBS, HRP conjugated anti-M13 antibody was added to the wells. The wells were washed after 45 min incubation and TMB substrate was added. The reaction was stopped after 15 minutes using 1N HCL and absorbance was read at 450nm.