Agrobacterium Causes Crown-Gall Disease?

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

The pGLO plasmid is engineered to express green fluorescent protein (GFP) in the presence of the sugar arabinose as well as the ampicillin resistance gene β-lactamase (bla) (Brown, 2011). Original E. coli HB101 do not have ampicillin resistance or the GFP gene allowing them to glow under UV light. In this experiment, we transformed E. coli HB101 with the pGLO plasmid by heat shock to make the bacterial cells competent, allowing the plasmid to enter the cell (Brown, 2011). The mixture of bacteria with pGLO plasmid were given recovery time after heat shock, then spread on LB/amp and LB/amp/ara agar plates. The bacteria mixture with no plasmid added were spread on LB and LB/amp agar plates and all four plates were incubated at 37°C for

The plasmid pGLO contains an antibiotic-resistance gene, ampR, and the GFP gene is regulated by the control region of the ara operon. Ampicillin is an antibiotic that kills E. coli, so if E. coli, so if E. coli cells contain the ampicillin-resistance gene, the cells can survive exposure to ampicillin since the ampicillin-resistance gene encodes an enzyme that inactivates the antibiotic. Thus, transformed E. coli cells containing ampicillin-resistance plasmids can easily be selected simply growing the bacteria in the presence of ampicillin-only the transformed cells survive. The ara control region regulates GFP expression by the addition of arabinose, so the GFP gene can be turned on and

Our world has changed dramatically since the day Antoine van Leeuwenhoek discovered microorganisms in 1676 using a simple microscope. In early days, scientists first thought life arose from inanimate materials. This theory, known as abiogenesis or spontaneous generation, was disproved later on by scientists including Lazarro Spallanzani and Louis Pasteur. The experiments conducted by these scientists showed that living things could only arise from preexisting life, or biogenesis. All life begins with a living cell, composing of five required components. These components are DNA, RNA, cell membrane, ribosome, and cytoplasm. As more investigations on bacteria were conducted, scientists were able to acquire a deeper knowledge of the microbiology and pathology of animals, plants, and humans.

The gallbladder and bile ducts along with bile juice perform significant functions, including digestion and fat absorption and excretion of various metabolites from the liver.

“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

The cat's gallbladder is a small, balloon-shaped organ that is located in the lobes of the liver. Its primary function is releasing bile into the digestive tract to aid in the digestion of food. Cholelithiasis is a condition that causes small stones, or choleliths, to form in the gallbladder. These stones are typically made of calcium carbonate mixed with other secreted substance and minerals.

Gallstones are described as digestive fluid that has hardened which can be very painful. They form in the gallbladder which is a pear shaped organ that is located on the right side of of the abdomen right underneath the liver. Gallstones can be many sizes which varies how much the pain is. They can be as small as a crumb to a size of a golf ball! Ouch! Also, some people have more than one, while other people may just have one. More than likely, you will have symptoms if you have gallstones and if you do have symptoms, most people have to get gallstone removal surgery. These symptoms may include, sudden pain in your upper right side of the abdomen or under the breast, back pain between shoulder blades, right shoulder pain, nausea, or vomiting.

The goal of this experiment was to investigate genetic transformation of E.coli through the reaction of organism to the vector pGLO plasmid. As mentioned, the pGLO plasmid contains genes coding for resistance to ampicillin (amp), and genes coding for production of the green fluorescent protein (GFP) which glows under UV light in the presence of arabinose (ara),which serves as a reporter gene. This green fluorescent

Gallstone disease represents an important issue in the healthcare system and one of the most common and costly of all digestive diseases if we consider the number of cholecystectomies, which are performed annually all over the world, and the hospital admission rate for complicated gallstone disease (Guarino et.al., 2013). Gallstone disease is a disorder where genetics and the environment plays a major factor. Some of the risk factors that contribute to the disease are age, gender, race, parity, and dietary factors (Guarino et. al., 2013). Gallstones are hard deposits that can be extremely small or as large as tangerine, and they tend to roam around in your gallbladder. Gallstones develop where there is too much cholesterol in the bile. When

The predicted antigenic regions were chimerically expressed in a bacterial system. Briefly, two sets of PCR primers were designed according to gD gene sequence available in GenBank (accession number: NC_001847) to amplify gene fragments encoding aa 20-160 (nucleotide 118953～119375, Δ gD1) and aa 257-344 (nucleotide 119664～119927, Δ gD2), respectively. The primers sequences for amplification of Δ gD1(P1 and P2) and Δ gD2 (P3 and P4) were listed in Table. 1.

In this last situation, resistant genes become embedded in small units of DNA, called transpons, which can easily move into other DNA molecules. Making matters worse, many bacteria have specialized transpons called integrons, which act like flypaper when catching new genes (3).

Abstract:Conjugation is a natural occurring process that involves the transfer of DNA from one cell into another through a physical connection between the cells. In the following experiment, two strains of Escherichia coli bacterial cells (donor F'lac+strs and recipient F-lac-strr) underwent conjugation to produce a transconjugant strain (F'lac+strr). MAC plates and streptomycin were utilized to determine if conjugation had occurred. When plated, the donor colonies appeared red and the recipient colonies appeared white. The transconjugant plates showed red and white colonies. Using alkaline lysis miniprep, a DNA plasmid was isolated from the donor and transconjugant strains and FIGE electrophoresis was used to determine the size of the

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 coding region of the gene is usually fused to a promoter, most commonly used is the 35S promoter from cauliflower mosaic virus (CMV), in order to promote higher expression levels. (Snow et. al, 1997) The popular method for genetic engineering of crop plants is natural gene transfer via an Agrobacterium tumefaciens vector, a bacterium normally found in soils. The transfer-DNA (T-DNA) vector is made by inserting the desired gene fragment in between specific 25bp repeat domains in the bacterium. The vector is then inserted into the Agrobacterium and "the virulence gene products of Agrobacterium actively recognize, excise, transport, and integrate the T-DNA region into the host plant genomes." (Conner et. al, 1999) The amount of DNA transferred is only about 10kb and the nature of the gene is usually well understood. The expression of the gene introduced can also be controlled by adding additional sequences that might allow the gene to be constitutively expressed, expressed only in certain cell types, or expressed as a result of different environmental changes. This method of gene transfer, however, will only work for the natural host range of the bacterium and therefore other methods are used for additional crop plants. Such methods are uptake of naked DNA by electroporation or particle gun bombardment. The use of genetic markers, as mentioned previously, allows for the preferential growth of cultures that contain the new genetic