Cloning and Sequencing a Portion of Glyceraldehyde-3-phospate Dehydrogenase Gene from the
Leaves of Soleirolia Soleirolii
December 13, 2013
Introduction
Without energy, plants cease to exist. Energy is a vital part in the process known as photosynthesis. In this pathway of photosynthesis, glycolysis helps yields ATP and NADH. The ATP released through glycolysis is energy that is reused to go through another production of photosynthesis. The Glyceraldehyde 3-phsophate dehydrogenase (GAPDH) gene is important in the process of glycolysis. Glycolysis is involved in energy production in carbohydrate metabolism. It helps breakdown glucose into pyruvate, thus releasing energy. GAPDH is located in the genome of plants and codes for an enzyme that catalyzes the process of glycolysis. The enzyme catalyzes NAD+, which produces an intermediate of 1,3-biphosphoglycerate (Niu et al, 1994). GAPDH is also involved in endocytosis in the cytoplasm of a plant cell. In the nucleus, it not only takes part in tRNA export but it is also involved in the translational level of gene expression through DNA replication and repair (Sirover, 1997).
In these series of lab experiments preformed throughout this project, the main objective was to isolate, sequence and clone GAPDH. The GAPDH will be extracted from a plant called soleirolia soleirolii, commonly known by it’s household name, baby tears. Another plant that will be looked at closely is Arabidopsis. This plant will act as the control throughout the experiment. The first step of this experiment is to extract the GAPDH gene from the plant. Column chromatography helps in separation and determination of compounds such as amino acids, sugars, phosphate esters and nucleotides (Thomas...
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In the light independent stage of photosynthesis ATP is again used to break down a molecule. In the Calvin cycle after glycerate 3-phosphate is reduced, then ATP breaks down and loses a phosphate group (becoming ADP). The phosphate group is then gained by the glycerate 3-phosphate molecule and it becomes triose phosphate. ATP is then used furthermore in product synthesis (anabolism) this is where energy is required to convert the triose phosphate into more complex molecules such as amino acids or lipids.
The majority of life on Earth depends on photosynthesis for food and oxygen. Photosynthesis is the conversion of carbon dioxide and water into carbohydrates and oxygen using the sun’s light energy (Campbell, 1996). This process consists of two parts the light reactions and the Calvin cycle (Campbell, 1996). During the light reactions is when the sun’s energy is converted into ATP and NADPH, which is chemical energy (Campbell, 1996). This process occurs in the chloroplasts of plants cell. Within the chloroplasts are multiple photosynthetic pigments that absorb light from the sun (Campbell, 1996).
ABSTRACT: Chloroplasts carry out photosynthetic processes to meet the metabolic demands of plant cells (Alberts, 2008). They consist of an inner thylakoid membrane and a stroma. (Parent et. al, 2008).In this experiment we demonstrate the unique protein compositions of isolated thylakoid and stromal fractions from broken and whole spinach chloroplasts. Because these compartments carry out different metabolic processes, we confirm our hypothesis that performing SDS-PAGE on these fractions will result in distinct patterns on the gels. In isolating and analyzing nucleic acid from broken, whole, and crude chloroplast samples we demonstrate that genes for photosynthetic protein psbA are found in chloroplast DNA, while genes for photosynthetic enzyme
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...e involved in the plants carbohydrate metabolism. This response causes the plant’s cell walls to be rearranged and strengthened. THis would increase the plants resistance to infection and the uptake of harmful chemicals.
All living things depend on a source of energy for their survival. These sources may vary from one species to another. For example, human’s and animal’s main source of energy is food, while plants main source of energy is sunlight. Plants lack the ability to move and look for sources of energy, runaway from predators, or avoid Abiotic stress. Instead, they have Photoreceptors such as chlorophyll found in plants’ chloroplasts which absorbs light and changes it into a cascade of electron transfer that serve as the main source of energy for plants. Moreover, different kinds of proteins regulate the plants life cycle such as phytochrome, cryptochromes, and phototropins. These proteins are mostly pigments that intercept light at different wavelengths and thus each photoreceptor is activated by different light conditions. Using their Photoreceptors, plants transform different light signals to regulate the plant’s growth, development, defense mechanisms, and stress responses. Most of the processes and mechanisms taking place in plant cells usually rely on signaling pathways. These signaling pathways depend on proteins that have different function in activating, inhibiting, or relying the signal from a protein to another. The most important proteins in these signaling pathways are kinases and phosphatases. Studies have recently shown important data that proves the interaction of these photoreceptors and some kinases and phosphatases, for example the interaction of phytochromes with PP2A phosphatases (Bissondial, 2005).
The two 3-carbon pyruvate molecules that were created from glycolysis are oxidized. One of the carbon bonds on the 3-carbon pyruvate molecule combines with oxygen to become carbon dioxide. The carbon dioxide leaves the 3-carbon pyruvate chain. The remaining 2-carbon molecules that are left over become acetyl coenzyme A. Simultaneously, NAD+ combines with hydrogen to become NADH. With the help of enzymes, phosphate joins with ADP to make and ATP molecule for each pyruvate. Enzymes also combine acetyl coenzyme A with a 4-carbon molecule called oxaloacetic acid to create a 6-carbon molecule called citric acid. The cycle continuously repeats, creating the byproduct of carbon dioxide. This carbon dioxide is exhaled by the organism into the atmosphere and is the necessary component needed to begin photosynthesis in autotrophs. When carbon is chemically removed from the citric acid, some energy is generated in the form of NAD+ and FAD. NAD+ and FAD combine with hydrogen and electrons from each pyruvate transforming them into NADH and FADH2. Each 3-carbon pyruvate molecule yields three NADH and one FADH2 per cycle. Within one cycle each glucose molecule can produce a total of six NADH and two
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Photosynthesis is a process in which plants and other organisms convert the light energy from the sun or any other source into chemical energy that can be released to fuel an organism’s activities. During this reaction, carbon dioxide and water are converted into glucose and oxygen. This process takes place in leaf cells which contain chloroplasts and the reaction requires light energy from the sun, which is absorbed by a green substance called chlorophyll. The plants absorb the water through their roots from the earth and carbon dioxide through their leaves.
When humans consume plants, the carbohydrates, lipids, and proteins that are broken down through two forms of cellular respiration. The two processes of cellular respiration displayed in humans are anaerobic or aerobic. The deciding process used depends on the presence of oxygen. Cellular respiration converts the material into useable energy called ATP. ATP is the energy form that cells can use to perform their various functions and it can also be stored for later use. Without plants, none of this energy could be produced and the herbivores that humans consume would not exist.
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