Recombinant Dna Technology As An Environment For Separation

This experiment was conducted as per the BCHM 310 Laboratory Manual [3]. The first objective of this experiment was to analyze the purity of the invertase fractions collected during experiment 6, and to determine the molecular weight of LDH-H4, LDH-M4 and invertase subunits. This was accomplished using sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS-PAGE). In this procedure, SDS, a negatively charged amphipathic molecule, was used to denature the proteins and to give each protein a similar charge-to-mass ratio [4]. As a result, most oligomeric proteins separated into individual subunits, and each subunit assumed a rod-like shape [4]. The distance travelled by each subunit, along the polyacrylamide gel, was a function of its molecular weight; where proteins with a greater molecular weight moved a smaller distance than proteins with lower weights [5]. Since SDS is not a reducing agent, and no other reducing agent was added, oligomers with disulfide bonds between subunits would have remained intact [4]. However, this was not expected to be problematic for analyzing invertase or LDH isozymes, as these proteins lack disulfide interactions between their subunits [2,6]. In addition, since invertase and LDH are homo-oligomers, each protein’s subunits were expected to migrate the same distance [2,6].

"Ethanol Precipitation of DNA and RNA: How it works." Bitesize Bio. N.p., n.d. Web. 20 Feb.

This technique separates Rubisco samples based on their size. The electrophoresis has a positive and a negative end. Positive charge proteins are loaded from the positive end and migrate towards the negative end. Negative charge proteins are loaded from the negative end and migrate towards the positive end (Sakthivel & Palani, 2016). The sample that contained the highest molecular weight of Rubisco will travel the shortest distance on the gel while the protein with the smallest molecular weight will travel the longest distance (Sakthivel & Palani, 2016). The size proportion of each Rubisco molecule correlates with the distance traveled. Rubisco will be in its purest form after running through SDS-page since each technique will increase the purity of the protein. If the salting out, the ion exchange and the SDS-page protein isolation techniques are performed on protein Rubisco, then it is purified and separated by solubility, charge, and size. The rationale of this experiment is to isolate the purest form of Rubisco so that it can perform carbon fixation at an optimal

Genetically modified organisms are created through the use of biotechnological methods, in which genes from one organism

The protein gel electrophoresis showed that the flow through fragment

Plasmids are small double stranded circular non chromosomal DNA molecules containing their own origin of replication. Hence, they are capable of replication independent of the chromosomal DNA in bacteria. Plasmids present in one or more copies per cell, can carry extra chromosomal DNA from one cell to another cell and serve as tools to clone and manipulate genes. Plasmids used exclusively for this purpose are known as vectors. The genes of interest can be inserted into these vector plasmids creating a recombinant plasmid. Recombinant plasmids can play a significant role in gene therapy, DNA vaccination, and drug delivery [Rapley, 2000].

The DNA being heated to 95° C. This will result in the weak hydrogen bonds that clutch the DNA strands together in a helix to break apart. This causes the separation of the strands resulting in the generation of single stranded DNA.

DNA, Deoxyribonucleic Acid, is the basic structure for all life, it is the blueprint, the instruction manual, on how to build a living organism. DNA is made up of four nitrogen bases, adenine, thymine, cytosine, and guanine which are connected by sugar-phosphate bonds. Through a process called Protein Synthesis, the nitrogen bases are the code for the creation of amino acids. Essentially, DNA makes amino acids, amino acids make proteins, proteins make organisms. This process has been taking place for much longer than scientists have been able to document. Those scientists are called geneticists and their field is genetics.

The testing of various proteins was performed by comparing the molecular weight of proteins using SDS PAGE. The molecular focus in the lab was the testing of proteins, which are macromolecules consisting of amino acid monomers linked through chemical bonds. These proteins have a hierarchy of structure that consists of folding that determines the direct function of each protein.. The molecular weight of these proteins were measured using SDS PAGE. SDS PAGE stands for sodium dodecylsulfate polyacrylamide gel electrophoresis. SDS is an anion detergent that binds with the protein structures and causes them to separate due to the change in bonding charge. SDS and heat are how the proteins are denatured. The process of denaturing a protein is breaking

A basic method in which we get specific genes integrated with another organism’s chromosome is as follows: Isolate the DNA from which selected gene is to be taken from and treat it with enzymes that will cut out that specific gene. These genes are then inserted into bacteria and grown into colonies being

Technology has flourished and has improved our way of living in many ways than one. In the medical world, it has definitely shaped our ways to helping patients through medication. For instance, diabetes, or growth and clotting disorders among the spectrum of human diseases related to protein absence or malfunction. A lot of the medications that we use today are created from bacteria, yeast, insect cells, and mammalian cells, which boost the production of the protein within the human body. For example, insulin is a very in demand medication due to the increase of diabetes as a disease. Before technology, insulin would

The crude protein extraction of E. huxleyi cells that were either grown in 23℃ or 18℃, was prepared by grinding the cells in liquid nitrogen until it was fine powder. The protein extract was denatured and solubilized by adding 800 μL of an extraction buffer (30mM Tris-HCl, 7 M urea, 2 M thriourea, and 4% CHAPS), 8 μL of a protease inhibitor cocktail, 16 μL of B-mercaptoethanol, and shaking it for half an hour at room temperature. This process was done in order for no contamination to occur because, this algae’s electrophoretic separation of proteins concentrations is low.   Then the extract was transferred to a 1.5 ml microfuge tube and it was centrifuged for 30 minutes at 15,000 xg at 4 °C. The liquid portion of the extract was transferred

The product that will result by genetically modifying this organism’s genome is known as a recombinant protein. The process will combine the human protein with a vector, a specific DNA molecule, which will transport the genetic material. This procedure will use a bacterial plasmid. The plasmid is

Recombinant DNA technology is also known as, DNA cloning, molecular cloning, and gene cloning. In this process, the DNA fragment that is from one organism is transferred into a self-replicating genetic element. Bacteria plasmids make perfect hosts for this process. In order to make a clone of a gene, a DNA fragment that has the gene of interest must be isolated form the chromosomal DNA by using restriction enzymes. Next the gene is then united with a plasmid that has also been cut with the same restriction enzymes. After the fragment has been joined with its cloning vector it is now known as “recombinant DNA molecule”. Once the molecule has been introduced into suitable hosts cells, the recombinant can the be reproduced along with the host cell DNA. This technology has been around since the 1970’s, and it can be used for many different reasons. But lately, scientists that have been working the Human Genome Project have been using it. They use this process so that they are able to copy genes and other pieces of chromosomes to generate

There are many ways that biotechnology can be achieved, one of these is Recombinant DNA technology, this has reached a stage where scientists can take a piece of DNA containing one or more specific gene from nearly any organism, including plants, animals, bacteria, or viruses, and introduce it into a specific crop species. This is frequently been referred to as genetic engineering (Persley, Gabrielle pg. 1). Another common type of DNA technology is genetically-modified organism (GMO). This is an organism that has been modified or transformed using modern techniques of genetic exchange.