Identifying the Genetic Basis of Microbial Iodate Respiration in Marine Environments Dissimilatory (respiratory) metabolisms provide microbes with energy from the reduction of inorganic oxyanions (e.g., NO3-, ClO3-) via the electron transport chain1. Iodate (IO3-) is an example of a thermodynamically stable oxyanion with oxidizing and bioenergetic properties comparable to those of oxygen. Iodate serves as an alternative terminal electron acceptor in the biotic dissimilatory iodate reduction (DIR) to iodide (I-), a process which plays a significant role in marine iodine cycling2,3,4. DIR remains poorly characterized, especially since initial research suggested that iodate reduction only occurs abiotically via photolysis or nitrate …show more content…
I will align the genomes of PST-1 and two related Pseudomonas species, P. chloritidismutans and P. stutzeri (all > 98.8% genetic identity5) and identify which loci are unique to PST-1. I will transform these loci into either the nitrate reducing P. stutzeri or chloritidismutans. For unique regions in PST-1 stretching across several kilobases, I plan to use large cloning vectors such as cosmids. I hypothesize that PST-1 will tolerate higher concentrations of iodate compared to the other Pseudomonas, and upon expressing the PST-1 DIR gene(s) in P. stutzeri or, P. chloritidismutans, I expect increased resistance to iodate along with the iodate reduction phenotype. I will ensure that the transformants are showing increased tolerance to iodate by measuring the conversion of IO3- to I- with ion chromatography7
Aim 2: Identify PST-1 Genes Involved in DIR I propose a forward genetics approach to identify the primary, auxiliary, and regulatory genes involved in iodate reduction. Sequencing and annotation will provide a reference genome for a random barcode transposon mutagenesis with next-generation sequencing (RB-TnSeq) experiment. This method, used previously in P. stutzeri to assess growth defects under different growth conditions8, uses unique bar-coded transposons to identify disrupted genomic loci (see figure below). Mutants will grow together under control conditions, be transferred
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 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
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
For this experiment, E. coli was best for genetic engineering because of their size, and their fast reproduction (Spilios, 2017). E. coli will be genetically transformed using an engineered plasmid. A plasmid is a circular piece of DNA which independently replicates and multiplies because it has its own origin of replication (Spilios, 2017). The pGLO is the plasmid used in this experiment. Plasmids are used as vectors and they contain manipulated genes such as genes coding for antibiotic resistance for drugs like ampicillin. This antibiotic resistance of such serves as the selectable marker in genetic transformation and for genetic transformation to proceed, the cell must reach competency which is the physiological state that is required for the vector plasmid to get into the cell for transformation (Spilios, 2017). While competency can be reached naturally in some organism, it must be reached artificially in E. coli through treatment with CaCl2 and exposing them to heat shock using incubation (Spilios, 2017).
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.
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.
22) Refer to Table 8.2. If the sequence of amino acids encoded by a strand of DNA is
Being a nurse may have their own up and downs but some strengths that I have to make it through to being a nurse is communication skills with patients or either customers. I can give good service and be someone they can talk to comfortably and being friendly with them. Another thing that I have is respectfulness because being a nurse you have to respect your patients no matter their race, color, and culture. You have to be a person to respect them so they see how determine you are as a person. Leading up to having determination to work as a nurse and to be someone that can help others as they can in society. I do have a strong determination to being a nurse since I thought about my career and I do want to succeed and graduate with it. Connecting
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
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.
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).
The Compromise of 1850 included The Fugitive Slave Law, a law forcing non-slave owners in the free Northern states to return escaped slaves to their Southern masters and participate in a system they did not believe in. Jehlen notes the reaction to this cruel governmental act by stating that "[t]he nation's growing guilt and apprehension is tangible in the overwhelming response to Uncle Tom's Cabin" (386). It seems hard to believe that people could find no wrong in making it a law to return humans as if they were property. In fact, Stowe wrote her most famous work, Uncle Tom's Cabin, at a most opportune time; indeed, she wrote it in response to the passage of the
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
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.
There is a difference among instructing a class in which multiculturalism is the emphasis and joining a fundamental multicultural, comprehensive viewpoint into the classroom environment. Given that "there is no worldwide structure of a multiculturalism development that is faultless for attaining all objectives for all students" (Chamberlin, 2005, p. 26), discovering a method to shape a multicultural basis for sequences across the disciplines may be a better goal for faculty in higher education organizations. Multiculturalism is an idea that cannot be overlooked in today's culture. It is actual, it is connected to the globalization of higher education, and it is going to do nothing but produce in wealth in the future of higher education locations in the United States. Therefore, "it is dangerous that universities and colleges play a leading function in arranging its elements to purpose successfully in a more pluralistic society" (Almarza, 2005, p. 1). With that said, this essay will discuss the pros and cons of teaching multiculturalism in college classrooms.