Bio Case Study

This experiment will be done as the first step in determining an unknown protein. The second part of the protein determination will be done using the Western blot technique. We will use the Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) method to separate proteins by their molecular weight. This method is often used to determine the purity and weight of a protein. Electrophoresis is a method of separating proteins based on their chemical and physical properties. Gel electrophoresis has a gel support mediums that includes polyacrylamide, starch and agarose. The gel is chemically inert, it is easy to handle, and can me made to fit a desirable porosity. This makes it good to use for molecules with high molecular weights. After separating the gel, it will be stained with Coomassie Blue to visualize the protein.

The role of prions in BSE is to trigger proteins in the brain to fold abnormally. These prions are formed by abnormally folded protein that causes neurodegenerative conditions, similar to that of Alzheimer’s disease. Normal prion proteins(PrP) are bound to the surface of the neuron and can be altered by infectious prions to become misfolded and take on a different conformation. These misfolded proteins tend to clump together, or aggregate, because of their shape. The altered proteins (PrPsc) then enter a normal brain and binds to the normal protein prions. This process continues to take place and affect the brain. Prions occur naturally in our bodies so this doesn’t stimulate an immune response allowing this conformation to continue to take place. This disease has no known cause but is generally associated with the ingestion of meat from cows who already have BSE.

There is now strong scientific suspicion that the agent responsible for the outbreak of prion disease in cows, bovine spongiform encephalopathy (BSE or 'mad cow' disease), is the same agent responsible for the outbreak of vCJD in humans. Variant CJD (vCJD) is not the same disease as classic CJD It has different clinical and pathologic characteristics. Each disease also has a particular genetic profile of the prion protein gene. Both disorders are fatal brain diseases with unusually long periods measured in years, and are caused by a transmissible agent called a

A balanced scorecard has four perspectives, Customer perspective, internal business perspective, Innovation and learning perspective, and financial perspectives. By using balanced scorecards, BIOCO knows what the customers needed is. It helps BIOCO to fulfill customers’ need better. For internal business perspective part, BIOCO can clear the corporate vision and strategy. BIOCO can adjust the strategy according to the pervious balanced scorecards. Also the balanced scorecards link everyone in the company and enhance the communications between each other. Meanwhile, Scorecards detailed the company’s business goals to everyone and guide every

Throughout this book are very interesting things to learn about prions and their effect on both livestock and humans. The perspective of many different scientists who tackled the problem was also shown. This book includes their successes their failures and now it is time to analyze the all of the information. What made these diseases, created by malicious malformed proteins, become such timeless pandemic? How are they transmitted? How is it possible for them to jump between species?! As the book shows, these questions are not easily answered but at least there are the brilliant minds of the scientists like Gajdusek and Prusiner to assist those who would analyze the information on prions.

Kuru was first seen in the mid-twentieth century and was at its peak in the late 1950s, due to the Fore tribe in Papua New Guinea having cannibalistic rituals for their dead. Every year 2% of their population would be killed off by the disease, which eventually led their tribe to the brink of extinction. It is thought that Kuru began when a Fore tribe member sporadically had developed Creutzfeldt-Jakob disease (CJD) and was later consumed after passing away. Creutzfeldt-Jakob disease (CJD) is a prion disease that spontaneously strikes about one person in a million each year (Deng, 2015). Prions are misfolded proteins that have the ability to make other proteins in the body misfolded, therefore being infected with one can start a surge of proteins being converted into more prion protiens (Minikel & Vallabh, 2013). Those who consumed

Prions are infectious protein agents that cause aggregation of the peptide PrPC and form large amyloid fibrils. These fibrils resist proteoysis and can have a detrimental effect of the functionality of cells and ultimately cause their death. (3). Misfolded PrPC will form stable aggregates denoted as PrPSc. PrPC is a soluble protein that is present in all organism whose function is still not totally clear (3). When PrPc converts to PrPSc there is an increase in the number of β sheets in the protein, which can allow for interaction and stacking of other misfolded PrPSc proteins, causing an amyloid fibre to form (5).

Proteins are substantial biomolecules, or macromolecules, comprising of one or all the more long chains of amino corrosive buildups. Proteins perform an endless cluster of capacities inside of living life forms, including catalyzing metabolic responses, DNA replication, reacting to jolts, and transporting particles starting with one area then onto the next. Proteins vary from each other essentially in their arrangement of amino acids, which is managed by the nucleotide grouping of their qualities, and which typically brings about protein collapsing into a particular three-dimensional structure that decides its activity.A straight chain of amino corrosive buildups is known as a polypeptide. A protein contains no less than one long polypeptide.

Depending on different disease types, clinical symptoms of prion diseases vary, however, they share similar histopathological features, including neuronal loss, astrocytic activation, spongiform degeneration, and prion protein aggregation (Aguzzi et al, 2008a;

On our SDS-PAGE, our protein standards were separated alongside our protein samples which allowed determination of the sizes of our protein in our sample. SDS denature proteins and coats them with a negative charge allowing for their electrophoretic separation by polyacrylamide gel electrophoresis (PAGE). As you can see, after being stained with the coomassie blue, we could detect proteins of various sizes and our unknown protein expressing GST was detected in our Pure B sample (Figure 2). The distance travelled from the wells on the SDS-PAGES by our protein standards were measured and compared to our protein ladder in Daltons (Table 2), from which we could construct our semi log plot. On the semi-log plot, our unknown protein within our Pure B travelled 27mm from the well with a size of 29,000 Dalton (Figure 3). To identify the specific protein in the gel, the western blotting method was used to detect which of the cultures (A or B) contains GST (Figure 4). After the cassette was taken out of the electrophoresis unit, colors of our protein standards appeared on top of our membrane surface. After blocking, primary antibody incubation, and secondary antibody incubation, we could clearly identify our GST in our Pure B and Crude B (Figure

These can then infect other PrP molecules, causing them to misfold, thereby propagating the loss of function. Prions have the distinctive ability to form sticky patches on the brain, eventually killing off healthy cells. The authors of the study recommend extreme caution and stringent safety protocols when dealing with patients with prion disorders. Since proteins, unlike viruses or bacteria, cannot be disinfected by common sterilization regimens, usage of different surgical equipment for prion diseases and other neurodegenerative conditions is highly

since all unbounded protein removed and only bounded protein (PTEN and other protein that has affinity toward the ligand) left. Note that, some samples diluted to fall in the standard range and there are samples excluded for not being in the range. The eluted sample should include a purified PTEN and the SDS-PAGE in figure 4 confirm these expectations. The red arrow refer to PTEN and it has the same Mw 63.1 as the pervious gel, which indicated that this band is PTEN band. The eluted samples present in the SDS-PAGE shows other bands with PTEN, which represents other proteins that have affinity toward the ligand. Other purifying technique can be add to this protocol such as size exclusion chromatography to obtain more purified sample. In addition, mass spectrometry can be used to obtain the sequence and the Mw of the band (after digestion the protein from the gel) to confirm that band represent PTEN

Prion Protein (PRNP) is encoded by a gene, which contains a highly unstable region of five tandem octapeptide repeats. An octapeptide repeat is a fragment from the protein which is composed of eight amino acids joined by peptide bonds, repeatedly copied along the amino acid chain. Within PRNP; these are tandem meaning that the octapeptide sequences are in such an arrangement that they are lined up one behind the other (Mastrianni, 2009).

P1NP is derived from extracellular processing of procollagen type I molecule. Procollagen contains amino- and carboxy-terminal extensions that are enzymatically

My vision is to understand at a mechanistic level the unique immunology associated with persistent herpesviruses such as cytomegalovirus (CMV), and to translate this knowledge into development of herpesvirus-based vaccines to control highly pathogenic emerging zoonotic viruses, either in the reservoir/spill-over animal species or human host. For greatest impact, I believe that science must not stop at the gain of basic scientific knowledge, but rather needs to continue through towards the exploitation of this knowledge to solve real-world problems. The structural determination of DNA would have remained only a fine example of the elegance of biological simplicity had it not been for the molecular biology explosion that it precipitated.