chinesis. A construct of R751::Tn4351 (the physical map of R751::Tn4351 and restriction sites are shown in fig. 7) was selected for introduction into F. chinesis to discover if the introduction and insertion of the vector R751 and the transposition of T4351 into the F. chinesis chromosome by a triparental mating occurred. One parent was E. coli GJ342 which carried a helper plasmid, the second parent was E. coli HB101 which contained R751::Tn4351 and the third parent was the F. chinesis target strain. 189 colonies were isolated on LB agar plates which in passage in fresh media were able to grow in 200µgml-1 erythromycin.
Introduction For two days, on the 14th and 15th of April, a field excursion to Hastings Point, New South Wales was conducted. At Hastings Point, topography, abiotic factors and organism distribution were measured and recorded, with the aim of drawing links between the abiotic factors of two ecosystems (rocky shore and sand dunes), the organisms which live in them, and the adaptations they have developed to cope with these conditions. Within these two ecosystems, multiple zones were identified and recorded, and this report also aims to identify the factors and organisms associated with each zone. Lastly, using data and observations from the past, predictions for the future of the rock pool ecosystem were made.
Hypothesis: If the pGLO plasmid is inserted into E. Coli bacteria, then the surviving cells with the arabinose will glow green under UV light as they will be genotypically identical and phenotypically different. Experimental Design: In this experiment, recombinant DNA technology was used to modify the genetic properties of an organism. First a plasmid was created which consisted of ori, araC, GFP, bla and pBAD promoter. The GFP is a jellyfish gene that codes for the production of green protein; bla breaks down ampicillin which then can be used to select only the bacteria containing the bla gene by placing ampicillin in the growth medium.
However, DNA can code for certain proteins, such as GFP that would cause the bacteria to glow after going through genetic transformation, explaining why the presence of arabinose sugar caused fluorescence. For this reason, it makes sense that the transformed bacteria is fluorescent, but the plasmid itself is
This time, we used minimal agar plate. The plate was labeled into two sections, Trsf and Mut. The Trsf colony on the LB plate was looped and spread on the Trsf section of the minimal agar plate and the Mut was done likewise. The plate was then incubated at 30℃ until the next course day. Only the wild-type Acinetobacter gene enables the bacteria to grow on minimal medium, therefore if growth was seen on the Trsf section then we would expect that the mutant had transformed and picked up DNA from its surroundings.
The plasmid and the gene are both cut using restriction enzymes. To incorporate the gene into the plasmid, both the plasmid and gen stuck together by a bacterial enzyme called ligase. The plasmid with the foreign gene can then be inserted into the bacteria One specific plasmid is pUB110 which is circular whose host is Bacillus subtilis. It has a plasmid size 2.3 kpb and copy number between 20 - 50. We can selectively grow bacteria that contain this plasmid by using selective and differential media.
Without this antibiotic, every single plate would have lots of bacteria on it, only the one successful plate(+pGLO LB/amp/ara) would have a few glowing spots surrounded by lots of untransformed bacteria. The reason that this place was successful is because the transformed bacteria were able to resist the antibiotics, and also were also able to use the gene to make fluorescent proteins because the sugar arabinose was
The process of transformation is used often to evolve or change a cell by implanting new genes which can safely alter the genes already located in the organism we are trying to change. In this lab, the goal was to transform E coli. to glow in the dark, or in other words to make transform it and give it the ability to be bioluminescent. Bioluminescence is one of nature’s most useful and stylish abilities. In many species the ability to create a glow based off the DNA located in the cell is used in a variety of ways-
Cyanobacteria exist in many environments, which include terrestrial, freshwater, and marine habitats. They play a vital role as carbon dioxide consumers and oxygen producers in oceans. A recent study on Synechocystis, a species of cyanobacteria, has shown that these bacteria have the ability to actually see light. This is vital to the metabolism of the Synechocystis because they need light from the sun to produce energy. Synechocystis' bodies functions as lenses.
However, when these bacteria are grouped together to have high cell density, the molecules they secrete amount to a certain number, and once that number is reached, the behavior of that bacteria is switched on and in this case, bioluminescence is created. Similarly, in my project, I am screening the anti bacterial activity using oils. Before I use the oil, I have to culture the bacteria overnight so that I could use them in the plates after 16-18 hours of incubation. Based on the talk, I believe that I have an idea on how bacteria grow. This Ted Talk has inspired me about science in numerous aspects.
Introduction Bioluminescence is the emission of light by living organisms arising by exergonic chemical reactions. The term ‘bioluminescence’ originates from the Greek bios for "living" and the Latin lumen for “cold light" emission as less than 20% of the light generates thermal radiation. This has been reported in many terrestrial and aquatic organisms including bacteria, fungi, insects, algae, squid etc. Some of the bioluminescent organisms occur in symbiotic relationship with the higher organisms. The enzyme that catalyze the bioluminescence reactions is called luciferase, and one of the component substrates is designated as luciferin.
1. Introduction of exogenous DNA into animal cell lines, plant protoplast, yeast protoplast and bacterial protoplast. 2. Electroporation can be used to increase efficiency of transformation or transfection of bacterial cells. 3.
Task 6 Produce an illustrated article for a magazine that examines E. coli structure, growth conditions and curves and how it colonises a new environment. (M2) Structure E. coli consists of an outer membrane which contains lipopolysaccharides, a periplasmic space with a peptidoglycan layer, and an inner cytoplasmic membrane. Some strains of bacteria are pileated and capable of accepting and transferring plasmid to and from other bacteria and such property enables the E. coli under stress conditions to survive. It can perform complicated metabolism to maintain its cell growth and cell division, even though it has extremely simple cell structure which has only one chromosomal DNA and a plasmid.
This is done in a test tube by cutting the gene apart with enzymes and replacing parts of
Garland Science. New York. Willey JM, Sherwood LM and Woolverton CJ. Microbiology. 2011.
