Each solution was then plated on an LB/kanamycin/IPTG plate. Additionally transformation #2 was plated on a LB/kanamycin plate, and transformation #4 was plated on an LB plate. The purpose of including kanamycin on the plates was to selectively screen for plasmid containing E. Coli cells. The IPTG was to selectively screen for E. Coli cells with the plasmid containing the EGFP insert. The plates were then incubated over night at 37° C. DNA Purification The objective of this experiment was to isolate plasmid DNA.
The secondary antibody would the overlap the primary antibody by binding to it. The secondary antibody (anti-rabbit HRP labelled) used is conjugated with an enzyme Horse radish peroxidase which then binds to its substrate tetramethyl-benzidine (TMB) to produce a blue color indicating the presence of lysozyme. SDS-PAGE gel electrophoresis is a process which separates proteins based on molecular weight. The purpose of this method is to separate out the lysozyme by its weight. The weight is known to be 14.6Da.
1972. The dye exclusion test for cell viability: Persistence of Differential staining following fixation. In Vitro 7(5): 323-329. V. Answer to Guide Questions 1. Other vital stains available for assessment of cell viability are safranin, aleian blue, eosin, erythrocin B, congo red, and nigrosin (Yip and Auersperg 1972).
Title: Genetic pGLO Transformation Introduction: Genetic transformation is a transformation that involves a change in genes. Transformation is the process by which the genetic material carried by a cell is altered by the incorporation of foreign exogenous DNA into its genome. In this lab, we used a procedure called heat shock, accompanied by a bacterial plasmid vector, to transform bacteria with a gene that codes for GFP (Green Fluorescent Protein). A vector is an agent “employed to transfer the gene from one organism to another” (Lab manual). Two common vectors are phages and plasmids.
1992) on the food vacuoles of the parasites. Parasites obtain amino acids for protein translation and formation of their cell membrane from digestion of hemoglobin in the host. The undigested free toxic heme groups form reactive oxygen species. The natural process to remove toxic heme group from the parasitic cell is by the bio crystallization of heme to haemozoin (Pandey AV et al. 2003).
Introduction A mutation is a heritable change that is passed from the mother cell to progeny cells. Mutations may lead to good, bad or neutral phenotypic changes in the organism. They may occur spontaneously as in random DNA replicative errors or may be induced by mutagenic chemicals or radiation. Besides mutations, another way that bacteria achieve gene diversity is through the three known mechanisms for intercellular gene transfer. They are transformation, a genetic process which free DNA is incorporated into a recipient cell, transduction, a process which bacterial virus transfers DNA to another cell, and conjugation, a form of horizontal gene transfer which requires cell-to-cell contact.
The caseinolytic protein protease (ClpP) is a highly conserved serine protease present in bacteria and higher organisms. ClpP is responsible for cell homeostasis and among other duties for the regulation of bacterial virulence in several pathogens including Staphylococcus aureus and Listeria monocytogenes. Significant interests in ClpP inactivation started with the discovery of its crucial role in virulence of these pathogens that cause severe infections in the clinics and are difficult to treat through the occurrence of multidrug resistance . There have been efforts to discover and develop small molecules that perturb the activities of ClpP.Stephan A. Sieber etc. demonstrated that selective inhibition of ClpP in Staphylococcus aureus resulted in a drastically decreased expression of major virulence factors.To date, β-lactone is the main inhibitor scaffold that exhibits specificity for ClpP, it is important to systematically analyze the structural features and expand the chemical space of putative inhibitors.
Pulse field electrophoresis is used for the separation of larger fragments of DNA. These fragments result from digesting a bacterial genome with a rare-cutting restriction enzyme. The pattern of DNA produced on the gel is used to differentiate different strains of bacteria. For the identification of individuals like humans or other organisms, Restriction fragment length polymorphism (RELP) analysis has turn out to be very practical. After isolation of DNA from the source it is digested enzymatically with the help of restriction endonucleases.
Enterococcus Faecium: A bacterium that is known to live inside our intestines. It is part of the Genus Enterococcus. Its form is round and spherical. Background: Aim/Purpose of the experiment: The aim of this experiment is for us to research and find about whether the Enterococcus Faecium bacterium has developed any immunity or resistance against the eight antibiotics that we will test in this experiment, to see whether they react at all and if yes, if they are reacting against the bacterium and are developing an immunity whatsoever, that helps to contain an outbreak of the said bacterium. Antibiotica that will probably work: