Identifying The Genetic Basis Of Microbial Iodate Respiration In Marine Biology Lab Report

This same procedure is repeated to transfer worms from the dpy-13 starter plate to the OP50 seeded plate labeled “dpy-13”. These plates are then both incubated upside down at 20˚C for 48 hours. This incubation gives the worms time to grow in their new environment.

The citric acid cycle has been described as “the hub of the metabolic wheel”. Discuss the roles of the citric acid cycle in the oxidation of various fuel molecules and the provision of carbon skeletons for biosynthesis.

The addition of 5-bromo-4-chloro-3-indoyl-β-thiogalactoside (X-gal) in the agar mix acts as an artifical substrate for the enzyme so it produces blue E.coli colonies when hydrolised by the β-galactosidase meaning those specific colonies do not contain the plasmid with the CIH-1 insert (non recombinant E.coli) (Coventry University 2016). Isopropyl-β-D-thiogalactoside (IPTG) an artifical transcription inducer of the lacZ gene is also added into the agar mix, binding to the repressor gene and inactivating it. Therefore when the CIH-1 gene is incorporated within the plasmid MCS and inserted into the E.coli

As is the case for almost every mechanism in an animal body, oxygen is required for metabolic processes. This process, aerobic respiration, is the production of cellular energy in the presence of oxygen. Due to this fact, the metabolic rate of crayfish can be studied and calculated based on the rate of oxygen consumption for the organism in question (Meade et al, 2002).

Oxidation-reduction reactions can be used to stereochemically control and produce many different organic molecules. The oxidation step in this process increases the number of carbon oxygen bonds by losing a hydrogen and breaking

One of the most imperative functions in maintaining the development of evolution is the frequency of genetic transformation: the injection of foreign DNA into another organism’s DNA. This term is defined by the actions of a vector, but more specifically by the actions of plasmids and phages. However, in this experiment we are primarily focused on the effect of the pGLO plasmid transformation of GFP on the E. coli bacteria by introducing a second chromosome or a plethora of cloned plasmids. (Bassiri 2011)

The field of biotechnology involves the concept of genetic engineering, altering the DNA/genetic material of an organism using information from a different one. The process in which bacteria can obtain this manipulated genetic information from another source is called genetic transformation. The goal of this experiment was to genetically transform Escherichia coli bacteria’s DNA by inserting the vector pGLO plasmid which codes for ampicillin resistance as well as the green fluorescent protein, GFP. For the experiment, the E. coli bacteria were separated into two groups; control and

The transposon in this experiment is contains kanR in between the inverted repeats on either end, which will be transposed from the plasmid pVJT128 to the chromosome of the recipient bacteria.

To start the experiment two small plastic tubes were labeled negative pGLO and positive pGLO as the first step in this experiment. A pipet was then used to move 250 microliters of calcium chloride into each tube and the tubes were iced. Bacteria was added using a new sterilized loop each time to the positive and negative pGLO tubes. The loop was twisted around in the tube to ensure that the bacteria was evenly mixed into the solutions and the tubes were iced once again. Another loop was dipped into the tube containing the plasmid and it was removed when there was a visible slight film of residue. The plasmid was dipped into the tube that was labeled positive for the pGLO gene. The two tubes labeled positive pGLO and negative pGLO were iced

22) Refer to Table 8.2. If the sequence of amino acids encoded by a strand of DNA is

Living systems biologically depend on the six major nutrient elements (C, H, N, O, S, and P) and complementing trace elements that can interchange for one another if they share chemical similarities. But, there are no prior reports of substitutions for the major nutrient elements. Wolfe-Simon et al. presents evidence that As can substitute for P in the biomolecules of bacterium. Although Since As is a chemical analog of P, downstream metabolic processes are usually not compatible with As-incorporating molecules due to differences in the reactivities of As and P compounds. But, Wolfe-Simon et al. hypothesized that AsO43- could substitute for PO43- in an organism with the ability to cope with the instability of AsO43- compounds. Wolfe-Simon et al. experimented with AsO43- combined with no added PO43-, to select for and isolate GFAJ-1 of Mono Lake. Their data (from ICP-MS, synchrotron x-rays, NanoSIMS, etc) showed As-dependent growth by GFAJ-1 that was accompanied by AsO43- uptake and assimilation into biomolecules (nucleic acids, proteins, and metabolites). By using As as a selective agent and excluding P, it was shown that GFAJ-1 is not an obligate arsenophile and grew much better when provided with P. Also, GFAJ-1 coped with the instability of AsO43- esters due to poly-β-hydroxybutyrate rich vacuole-like regions and possibly from intracellular regions or mechanisms that exclude water.

In order to achieve this goal they created variants of each gene. These genes were all removed by a similar process by which the pieces of the gene were cut with different restriction enzymes and purified and cloned in order to give us different plasmids. They were amplified using PCR and these were then integrated into the regions and then grown on media and observed. When there was excision of the gene they were looking to remove these strains were cultivated and given designations. Strain WR 441(hDHFR-2) has the hdrA was deleted, WR445 has a deletion of both genes and WR446 (hDHFR-1) has a deletion of hdrB, WR 447 had hts deleted from it. These strains in turn were grown on three different Medias HY which was the rich media, M which was the minimal media and agar plates. In these experiments they found that WR441 needs nothing else and grew on each media with no other enrichment. However WR446 and WR 445 could grow on minimal medium containing both trimethoprim and thymidine, but only when that medium was supplemented with glycine, methionine, pantothenic acid and hypoxanthine. Another strain WR447 was grown and found that it was auxotrophic for thymidine. However when plasmid pHE4 was added to it could grow on minimal medium containing trimethoprim and thymidine. When the hts was removed the hdrB gene did not have a strong promotor which appeared when the hDHFR-2 didn’t express itself enough to give the organism the trimethoprim resistance it need to get a high copy number. Because of this we can use the hdr gene as an expression point for Hf.volcanii. At the lowest level of expression, only complementation of purine, glycine, pantothenate and methionine autotrophy will occur When the gene has a higher level of expression we can see that there will be prototrophic growth in minimal medium, and when the gene is working at its highest level it will give a resistance to

Antibiotic resistance is and continues to be a global public health issue1.One of the main concerns stems from the ability for bacteria to obtain antibiotic resistance very easily as a result of chromosomal changes, or through plasmids and transposons which generate an exchange of genetic material2. Resistance can also result from single or multiple mutations1,3. To combat this rising force, scientists must research and analyse the many possibilities of mutations in a variety of genes and proteins in specific bacteria and ways to combat them.

Cellular respiration is a procedure that most living life forms experience to make and get chemical energy in the form of adenosine triphosphate (ATP). The energy is synthesized in three separate phases of cellular respiration: glycolysis, citrus extract cycle, and the electron transport chain. Glycolysis and the citric acid cycle are both anaerobic pathways because they do not bother with oxygen to form energy. The electron transport chain however, is aerobic due to its use of oxidative phosphorylation. Oxidative phosphorylation is the procedure in which ATP particles are created with the help of oxygen atoms (Campbell, 2009, p. 93). During which, organic food molecules are oxidized to synthesize ATP used to drive the metabolic reactions necessary to maintain the organism’s physical integrity and to support all its activities (Campbell, 2009, pp. 102-103).

Bacterial transformation is the process of moving genes from a living thing to another with the help of a plasmid.The plasmid is able to help replicate the chromosomes by themselves; laboratories use these to aid in gene multiplication. Bacterial transformation is relevant in everyday lives due to the fact that almost all plasmids carry a bacterial origin of replication and an antibiotic resistance gene(“Addgene: Protocol - How to Do a Bacterial