Genetically Bacteria And Its Potential Against Human Diseases And Disorders

“Bacterial illness is a result of complex interactions between bacteria and the host. During evolution, humans developed many ways to protect themselves against bacterial pathogens. On the other hand, bacteria have developed strategies to evade, subvert or circumvent these defenses” (Sousa, 2003) “One of the most important characteristics of bacterial pathogenicity is the various strategies developed by prokaryotic organisms to use host molecules for their own benefit” (Sousa, 2003). “To accomplish this, bacteria have evolved elaborate control mechanisms to turn genes on and off, varying the transcriptional activator or

difficile to treat host from this pathogen. They use C. difficile strains, plasmids and conjugal transfer system to conjugate C. difficile gene. C. difficile strain 630 and VPI 10463was used and they first grew it anaerobically in Brain Heart Infusion media or it can be grown on an OXOID agar with 0.5% yeast extract and 0.1% cysteine, BHIS (2013). They also grew the bacteria in modified HySoy Medium by fermentation of 4-81. They then designed synthetic TcdA and TcdB genes using double allelic substitutions in GT catalytic site residues and used 630 toxin genome sequence for the recombinant genes (Donald, et. al, 2013). After synthesis TcdA and TcdB genes with their full length was subcloned as 8.1 kb and 7.1 kb NdeI-BlII fragments into pMTL84123 vector and E.coli strain Stb12 was the recombinant plasmid used (Donald, et. al, 2013). PCR was done using 5’ and 3’ flanking ends of the sites NotI and NdeI RE. Toxin A and B mutants was further mutated to modify or revert the toxins by site-directed mutagenesis using internal 2.5 kb NdeI-HindIII or 3.3 kb NdeI-EcoNI fragment subclones (Donald, et. al, 2013). Clostridim difficile was then conjugated by conjugal transfer of plasmids from E.coli and centrifugation was done to harvest the bacteria (Donald, et. al, 2013). Transformation that appeared on the media or agar was purified onto a BHIS agar and tested genetically modified toxins (Donald, et. al, 2013).

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).

When reflecting back to experiment 3, Aseptic Technique and Culturing Microbes, I realized the large amount of microorganisms that can be found in everyday life. Many different types are found with in the human body. Theses experiments focused on two types of bacteria. First was Staphylococcus epidermidis, found on the skin, and second was Lactobacillus acidophilus, found in the gastrointestinal tract. Both have similar needs for growth when it comes to temperature, however, different growth environments are used.

Flow Diagram Homework Assignment Name: Preetam Ganti Using the table below, explain the purpose of each of the steps of the Experimental Flow Diagram (Figure 1 from your lab manual – it is provided on the next page) for answering the two research questions guiding this laboratory investigation: 1. Is the bacterial contamination at these three farms due to the same plasmid or different plasmids? 2. What is the frequency of tetracycline resistant bacteria in the beef farm cultures?

This experiment was performed to assess the efficacy of genetic transformations on bacteria via plasmid DNA coding for ampicillin resistance and green fluorescent protein. Genetic transformation was studied by taking transformed and untransformed Escherichia Coli (E. coli) and placing them on various media to observe gene expression via growth and color under UV light. The transformed E. coli were able to grow on ampicillin while the untransformed E. coli, which lacked the plasmid genes for ampicillin resistance, only grew on nutrient broth. In the presence of arabinose, the transformed E. coli glowed green. These results support the previous scientific understanding of bacterial competency, vectors, and gene expression and support gene transformations as an effective method to transfer the desirable DNA of one organism into another organism’s DNA. These results can be applied to real world issues such as medical treatments, food production, and environmental conservation.

Bacteria are the natural world 's unsung heroes. They receive bad rep due to the disease causing individuals like tuberculosis but the vast majority of bacteria on Earth are harmless if not beneficial to both the environment and humans. Take for example E. Coli a well known bacteria that lives within our intestines. This particular bacteria makes vitamins that we need in order to stay healthy. In this experiment we analyzed the changes seen in bacteria when adding to differing DNA plasmids; pUC18 and lux. To arrive to the most accurate results possible we had to have the E. Coli bacteria made permeable to the plasmids. So it was prepared with calcium chloride. One group of bacteria was given the pUC18 plasmid, and the other group was given the lux as well as the pUC18 plasmids. We then proceeded to incubate them in different containers. Some had ampicillin where as some had none. This was done in order to observe the effect of pUC18, and lux growth patterns. A control with neither plasmid was used to keep a basis for our experiments. Bacteria that had transformed and taken in the lux plasmid without ampicilin were able to illuminate, and only bacteria that gained the pUC18 were capable of surviving with ampicillin. The results observed reveal how foreign DNA can be adapted to fit into another cell 's DNA even incorporated to be an optimal part of the cell. This process makes it a very viable manner of being able to mass replicate advantageous genes, that would

Plasmid map of pRSETB. Primers were designed to amplify via PCR to cDNA. The PCR product was digested with XhoI and EcoRI enzymes and ligated into the pRSETB plasmid. The pRSETB plasmid contains a T7 promoter region, is ampicillin resistant, inducible with Isopropyl β-D-1-thiogalactopyranoside IPTG, a molecular mimic of a lactose metabolite that triggers transcription of the lac operon, and has XhoI and EcoRI cut sites. The pRSETB plasmid is transformed into dH5α E. coli and plated on carbenicillin plates. Colonies are selected and grown on a carbenicillin plate while PCR is used to check that the plasmid that was up-taken was not

While leading this analysis, gene expression was critical to the results of the non-transformed and changed cells. In its procedure of choice, the bacterial cells should have picked if they ought to allow the ampicillin safe quality, therefore indicating commendable components of quality expression. The Escherichia coli's cells were not changed, there seems to have been an error during the experiment. There should have been a slight change in a way that there should have been a colonial growth design in the bacterial cells that had ampicillin safe plasmid. (Hidalgo, Nuria Fernández) On the other hand the cells that were on the ampicillin (lux, control, and NP) demonstrated a colonial design because of the non-transformed cells dying due to

In addition, many of the drugs available to treat infectious disease actually originate from bacteria and fungi. In Biology, constant inquiry of the mechanisms of life lead to new discoveries every day. Clearly, microbes are very important for studying and understanding our world. Dr. Henkin stressed their importance in the seminar, specifically the study of bacteria and bacteria’s manipulation of gene expression.

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].

This essay reviews a basic microbiological research project that focused on identifying pathogenic strains of Escherichia coli (E. coli) from the poultry industry in a developing country such as Romania. The research methodology, results and its practical applications are discussed. 51 isolates of E. coli were taken from various samples of raw poultry meat and tested for eleven specific antimicrobial resistance genes by using different techniques. Many antimicrobial resistant strains were isolated from the meat samples. These strains were confirmed by polymerase chain reaction (PCR)-based method to identify the pathogenic DNA. It was further revealed that these E. coli had genes which produced proteins against many antimicrobials, making them resistant. Fortunately, the most pathogenic, Shiga-toxic gene was not detected in any of the E. coli found in poultry meat. This research was very important for Romania because it would help lay a foundation for future studies and research regarding poultry food safety and precaution in Romania.

The domestication of animals 30,000 years ago was the precursor to Genetic Engineering. Starting with the dog we have since been able to introduce desirable traits in all organisms. With the discovery of plasmids in the late 60’s we have been able to take genetic engineering even further. Plasmids are small circular DNA molecules used to amplify and replicate a gene of interest. These minute molecules have the ability to replicate with the chromosome or independently, allowing them to have up to a 100 copies in one cell. Plasmids are important because of their characteristics to transfer genes that occur naturally within them or acting as a vector to introduce foreign DNA into a host cell. With the use of plasmids genetic engineers are able to use bacterial transformation to make medicines. Bacterial transformation is the process in which a bacterial cell takes up foreign DNA and incorporates this DNA into its own. With the use bacterial transformation this part of Genetic Engineering has become the most important and widely used technique, creating life saving antibiotics and medicines. E.coli is generally used in these procedures because of its ability to adapt and grow exponentially. Genetic Engineers can then use large bacterial colonies, Ampicillin, and X-gal to indicate if b-glactosidase is present along with identifying the recombinant and non-recombinant colonies. The transformed bacteria that contains ampicillin will spread and survive either turning white

Biotechnology has a long history of helping human beings have a better quality of life. Over the decades, numerous scientists have made substantial contributions to biotechnology. Among these scientists, I think Alexander Fleming made the greatest contributions. In this paper, I will explain how Alexander Fleming made significant contribution to biotechnology and how his contributions have had a tremendous impact on our modern society. To explain these, this paper will specifically focus on describing three main points. These three points are: first, Fleming discovered the world’s first antibiotic called penicillin and the use of this drug has saved millions of people’s lives; second, penicillin has led to the discovery and development of many other antibiotics which are used today to effectively fight a large number of bacterial diseases; and third, the widespread use of penicillin and other antibiotics have increased the average human life expectancy significantly.

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