Method Experiment Results DNase footprinting Binding to a protein protects DNA. The DNA is first either radiolabeled or chemically marked and then exposed to DNase (or in situ generated OH radicals) and then analyzed by gel electrophoresis. The DNA target sequence can be determined. Varying the concentration leads to binding constants. In addition, the influence of activating or inhibiting chemical can be assessed.
In order to utilize casein, bacteria cells secrete proteolytic exoenzymes (amylases, proteases, pectinases, lipases, xylanases and cellulases) outside of the cell that hydrolyze the protein to amino acids. The amino acids can then be used by cells after crossing the cell membrane via transport proteins . Starch hydrolysis test is used to differentiate bacteria based on their ability to hydrolyze starch with the enzyme α-amylase or oligo-l, 6-glucosidase. These enzymes hydrolyze starch by breaking the glycosidic linkages between the sugar subunits. It aids in the differentiation of species from the genera Corynebacterium, Clostridium, Bacillus, Bacteroides, Fusobacterium and members of Enterococcus .
Purification of GFP from E.coli via Hydrophobic Interaction Chromatography Priscilla Mariel M. Cadiz Biology Department, De La Salle University, 2401 Taft Ave, Manila, Philippines *Email: firstname.lastname@example.org Protein purification is an important techniquie in order to study the function and structure of proteins. Protein purification involves the process of removing contaminants and isolating desired proteins. Chromatography is one of the methods used for purifying and isolating proteins. Green Fluorescent Protein (GFP) is the desired protein to be isolated from the bacteria Escherichia coli which allows the bacteria to fluoresce. GFP contains abundant hydrophobic sites and because of this, Hydrophobic Interaction Chromatography
Proteins can undergo the process of degrading by the proteasome or by lysosomes. Transfer of proteins to lysosomes for the process of degrading, or autophagy, can occur through dissimilar mechanisms. There are three vital natural processes of autophagy in the cell,
THE SEQUENCING AND ASSEMBLING OF THE WHOLE GENOME OF AN ORGANSIMISM The sequencing and assembling of the whole genome is designed to help people understand and visualize how long a strand of DNA can be constructed from smaller overlapping DNA sequence. The genome sequencing is a very important aspect in molecular genetics because it help and gives us an understanding on how a genome completely works, How genes combine together and direct growth, development and maintain the whole body of an organism. The body of an organism is so complex and more complicated but it helps to study the gene expression of a specific tissue or organs and most importantly to study the human variation, how humans are closely related to other organisms. There are
Disrupting cellular structure is required to release the proteins from the cell. Purification of proteins begins with homogenizing the tissues, then subsequent fractionation and purification of cellular constituents. In this experiment, the source of protein was sprouting seeds. They were homogenized using a sodium phosphate buffer, pH 7, in a blender. After filtration, it was centrifuged.
Summary Endoplasmic reticulum is a eukaryotic organelle that forms interconnected network of cisternae, vesicles and tubules within the cells[1,2]. There are two types of endoplasmic reticulum: rough and smooth endoplasmic reticulum. The rough endoplasmic reticulum is covered with ribosomes in its membrane, these ribosomes are the site of protein synthesis. The ribosome free endoplasmic reticulum also know as smooth endoplasmic reticulum, its functions including lipid synthesis, drug detoxification and regulation of calcium concentration[2,3,4]. Furthermore, the endoplasmic reticulum can be isolated from animal soft by centrifugation method and the production form isolation can be used to study the metabolism of lipid and the recovery
Inside the cell, ara-C rapidly gets activated by many phosphorylation steps to form ara-CTP (cytosine arabinoside triphosphate). When this ara-CTP is incorporated into DNA/RNA, it inhibits DNA and RNA synthesis and triggers cell death. Thus DNA replication for mitosis is affected and the cells cannot divide rapidly. The first phosphorylation step is carried out by the rate-limiting enzyme Deoxycytidine
These protease enzymes will catalyze the breakdown of the protein into amino acids throughout the stomach and small intestines. Once broken down into amino acids, the microvilli will absorb them and will send them into our cells. Inside the nucleus of the cell, a DNA molecule will be unjointed by a RNA polymerase enzyme (brought by proteins), starting a process called transcription. This process allows RNA molecules to copy the genetic information from the unjointed DNA. The RNA polymerase begins by binding itself to the first base of a gene then it starts to copy the genetic information to a RNA molecule called a Messenger RNA (MRNA).
CRITICAL READING / A. THE KOJI PHASE Enzyme-substrate complex: Chemically, enzymes are protein molecules containing a specific area called active site. When a substrate comes and binds to this active site, biological changes takes place. The enzyme – substrate complex mechanism can be explained by different hypothesis. The lock and key hypothesis states that, the substrate fits exactly into the active site of an enzyme which is responsible for biological events to take place.