meant to prove that through genetic transfer using plasmid DNA‚ the E. coli can become bioluminescent and immune to the ampicillin. By adding plasmid DNA to the E. coli cells‚ the genetic composition of the cells will be different. I predict that the E. coli cells containing no ampicillin will be able to grow colonies. I also predict that the plates with plasmid DNA will show signs of bioluminescence. The plate with ampicillin present with no plasmid DNA will not be able to grow colonies and will not
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PA 17055 Page 2 ABSTRACT This study investigates gene regulation and how environmental arabinose and/or glucose can interact with genotype to influence phenotypic expression of the Green Fluorescent Protein (GFP) gene inserted in the plasmid under the control of a promoter called the pBAD promoter. Bacterial cells are a common choice for in vivo replication of DNA of interest‚ and in
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utilization of a strain of closely related bacteria controls and limits its damage‚ but it is also useful as a genetic engineering tool in plants. It is famous for taking advantage of its host by injecting DNA derived from its Ti (tumor inducing) plasmid into its host‚ causing the plant to create galls which excrete opines that the bacteria use as an energy source. A. tumefaciens have emerged as an important molecular tool for manipulating plants and creating genetically modified crops for research
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Lecture 10 Gene cloning BIOLOGY Campbell‚ Reece and Mitchell Chapter 20 Gene cloning technology Also known as: Genetic engineering or Genetic manipulation (GM) technology – implies precision engineering being applied to DNA molecules Recombinant DNA technology - implies that new combinations of DNA molecules can be made I.e. “recombinant” DNA molecules Overview of Genetic Engineering procedure 1. Making recombinant DNA molecules that can replicate in bacterial cells Genetic engineering
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environment) [Fred Griffith transformation did this]. a. Many have Calcium driven DNA pumps. (especially small DNA molecules) b. Electroporation – shock DNA to open pores in plasma membrane so DNA can enter cell c. Have small DNA molecules w/ 0 plasmids 1 ori and carry accessory genes 3) Restriction endonucleases attack DNA molecule from inside. Exonucleases – chop off 1 nucleotide at a time at the ends of the DNA molecule. Not in abundant in prokaryotes Bacterial DNA gets methylated – restriction
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foreign DNA from its surroundings. This DNA is then incorporated into the host cell’s own DNA. This transformation usually occurs within plasmids‚ small circular DNA molecules separate from its chromosome. After the recipient cell is infected with the DNA‚ the cell will move on with replication‚ producing offspring with traits encoded by the plasmid. These plasmids may replicate with the chromosome‚ or independently. This is how diseases are commonly spread‚ since one little bit of DNA can affect the
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ribosomes and other resources in the cell Plasmids Circular DNA Extrachromosomal NOT part of the E. coli genome “extra” DNA Contain a few non-essential genes Can give the bacteria additional “traits” Depends on the genes on the plasmid Can be exchanged between bacteria Recombinant plasmids Plasmids can be modified in biological labs Modified plasmid = Recombinant plasmid Plasmids can be used as cloning vectors to get the recombinant plasmid into E. coli Cloning vectors = way to get
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determining if any genetic transformation has occurred. By combining +pGLO LB and ampicillin we should get an ampicillin resistant gene and by using –pGLO we should create a non-genetic resistant bacteria. The pGLO plasmid has the GFP (green fluorescent protein) gene and the gene that allows the plasmid to be resistant to the antibiotic ampicillin. The most important part of this experiment is the “heat shock treatment” because the E. coli membrane becomes permeable and increases the competency of the cells
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have the COX1 gene‚ indicating that the probe was too specific. This could mean that the COX1 gene collected from the plasmid is different than the one from the potato‚ or that some other gene was labeled instead of the COX1 gene. Most likely the probe was specific enough that it could bind to the uncut plasmid DNA‚ but not to the mitochondrial DNA‚ or the DNA. The uncut plasmid DNA sample was given to us by another team working in the laboratory (Mitchell and Gytis)‚ that should not affect anything
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plant transformation method. The whole process lasted for over a period of 11 weeks and we were successful in getting transformed plants. Agrobacterium tumefaciens strain containing pMP90 (Ti-helper) plasmid and pCAMBIA1391 (T-DNA) plasmid was used for this plant transformation. pCAMBIA1391 plasmid was constructed by cloning Brassica rapa seed specific Napin promoter to control expression of GUS (beta-glucuronidase) reporter gene. Leaves were cut in small portions and were transferred to media containing
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