Learning Goals: Insert your uncut unknown plasmid into chemically competent DH-5 E.coli cells and use antibiotic resistance to confirm the success of the transformation. You should familiarize yourself with the various methods of transformation and the advantages/disadvantages of each type. You should also understand how heat shock transformation works and how chemically competent cells make this type of transformation possible. For this transformation antibiotic markers associated with foreign pieces of DNA will be used to help verify that the DNA of interest was successfully inserted into the vector.…
7) In gene cloning, the bacterial cells take up the recombinant plasmid DNA through a process called transformation. Bacterial cells can be transformed using electric pulsation or heat. The short electric pulse or a brief rise in temperature causes openings in the plasma membrane. The bacterial cells make copies of the recombinant plasmid DNA during cell…
A plasmid is a circular segment of DNA with an origin of replication. It can be duplicated and serve as an adjunct to the main bacterial chromosome.…
Plasmids are small circular autonomously replicating pieces of DNA that can be found inside of a prokaryotic bacterial cell. By barrowing a cell’s polymerase they replicate their own DNA. They are easy to extract from the bacterial cells due to their size. Plasmids are helpful for cloning foreign genes because of their ability to express antibiotic resistance as well their ability to be modified to express proteins of interest. A pGLO plasmid contains genes for the green florescent protein (GFP) as well as the gene for ampicillin resistance known as beta-lactamase. It also contains a gene regulation system (operon) that has the ability to control expression of the GFP gene in transformed cells known as araC. The source of GFP is naturally founds within a…
Genetic transformation is a process that primarily is inserting new DNA into an organism to change that organism’s trait. This process has many useful benefits when used correctly in different organisms. In this lab, bacteria was transformed by inserting DNA for Green Fluorescent Proteins. The DNA for these proteins were taken from bioluminescent jellyfish Aequorea victoria. One of the main lessons of the lab is learning of the use of ‘plasmids’. Plasmids are small pieces of DNA that usually code for one trait and are easily transferable between bacteria. This transfer of plasmids between bacteria is actually extremely helpful for them and are key in their survival. The plasmid that codes for the Green Fluorescent Proteins is accompanied with a gene for resistance to the antibiotic ampicillin. To ‘switch on’ the gene for fluorescence caused by the proteins, sugar arabinose must be added to the bacteria’s environment. If there is no sugar arabinose introduced to the plates, then the bacteria will appear white and will not glow, even if the gene for the proteins is successfully inserted. If the gene was successfully inserted and there is sugar arabinose present then the bacteria will glow a fluorescent green. The objectives for this lab is was to see the effects on bacteria in four different cases. The first case is the effect on bacteria when the gene for pGLO is introduced with LB (a ‘broth’ like substance that bacteria feed off of) and ampacillin. The second case is the effect on bacteria when the gene for pGLO is introduced with LB, ampacillin, and sugar arabinose. The third case is the effect on bacteria when no gene for pGLO is introduced, but LB and ampacillin is still introduced, The fourth case is the effect on bacteria when no gene for pGLO is introduced, but bacteria is still placed in a LB enriched environment. The…
b. In generalized_ transduction, all bacterial DNA has equal probability of being packaged into progeny phage…
Bacteria are able take in material though the cell membrane, resulting in a change in its phenotype. In transformation, the foreign DNA crosses through the permeable cell membrane with energy from various enzymes. In the E. coli bacteria cell, protein synthesis occurred, with these new genetic traits from the plasmid. In protein synthesis, there are three steps: transcription, RNA processing, and translation. In transcription, the enzyme RNA polymerase unzips doubled stranded DNA, and then attaches the corresponding nucleotides to the DNA sequence. Then, RNA Processing occurs, where the new messenger RNA strand is edited by a spliceosome. The spliceosome cuts out specific sections of the strand, and then the strand is able to come back together with all of its important parts. Unnecessary parts of the strand are called introns, while the important parts are called exons. Lastly, in translation, the mRNA is decoded by a ribosome to produce an amino acid chain. This amino acid chain, or polypeptide, will then start to fold into a protein. Now, after transformation, the foreign DNA from the P Vib affected the proteins in the E. coli bacteria, so that they now contain the genes for ampicillin…
Transformation is one of three processes for horizontal gene transfer by which genetic material passes from bacterium to another. “It is the acting of altering a genetic cell resulting from putting together exogenous genetic material from its surroundings through the cell membrane(s),”(Wikipedia, 2017, p.1).…
Abstract: In this lab, you will find out how plant and animal cells are alike and how they are different.…
A chromosome is necessary so that there is a “map” of the cells function and structure, cytoplasm is necessary as it is where most of the cell’s activity occurs, ribosomes are essential as they are the site of protein synthesis, which powers the activities of the cells, and the cell membrane is needed to give the cell structure and protection. Most cells have other structures based upon their function, and these extra structures are necessary in order for that cell to perform it’s function.…
Transformation is the genetic alteration of a cell resulting from the direct uptake of exogenous DNA from its surroundings. By doing this lab, we will answer the essential question: What influences transformation efficiency? Although there are many possible influences, our hypothesis is that plasmid has a positive influence. To test this hypothesis, we will set up an experiment in which we predict that if plasmid is present, then the transformation efficiency will increase.…
Chapter 8 The cellular Basis of Reproduction Inheritance Rain Forest Rescue • Scientists in Hawaii are attempting to "rescue" endangered species from extinction by promoting reproduction • Reproduction is one phase of an organism's life cycle – Sexual reproduction • Fertilization of sperm and egg produces Offspring – Asexual reproduction • Offspring are produced by a single parent, without the participation of sperm and egg • Cell division is at the heart of organismal reproduction -CONNECTIONS BETWEEN CELL DIVISION AND REPRODUCTIOn 8.1 Like begets like, more or less • Asexual reproduction – Chromosomes are duplicated and cell divides – Each daughter cell is genetically identical to the parent and the other daughter • Sexual reproduction – Each offspring inherits a unique combination of genes from both parents – Offspring can show great variation 8.2 Cells arise only from preexisting cells • "Every cell from a cell" is at the heart of the perpetuation of life – Can reproduce an entire unicellular organism – Is the basis of sperm and egg formation – Allows for development from a single fertilized egg to an adult organism – Functions in an organism's renewal and Repair 8.3 Prokaryotes reproduce by binary fission • Prokaryotic cells reproduce asexually by a type of cell division called binary fission – Genes are on one circular DNA molecule – The cell replicates its single chromosome – The chromosome copies move apart – The cell elongates – The plasma membrane grows inward, dividing the parent into two daughter cells THE EUKARYOTIC CELL CYCLE AND MITOSIS 8.4 The large, complex chromosomes of eukaryotes duplicate with each cell division • Eukaryotic genes – Many more…
8. Name the phase of the cell cycle in which cells spend most of their time doing their job.…
Transformation: in Eukaryotic cells: in humans means that difference in growth characters of cells, for example, the cells get transformed to cancer cells…
There are two different types of cells, there are prokaryotes and eukaryotes. Within eukaryotes there are different structures and similar structures. For example; in a plant cell they have a nucleus, mitochondria an ER, and a Golgi body. These are the same as animal cells; however they differ because plants cells have a cellulose cell wall, chloroplasts, large central vacuole and they use starch for storage whereas an animal cell has lysosomes, rough ER, smooth ER and ribosomes. Eukaryotes differ from prokaryotes because prokaryotes have no nucleus, they just have a loop of DNA and they also have no membrane-bound organelles. Prokaryotes also have a cell wall, along with a slime capsule, flagellae and plasmids.…