Bacteria are the only organism capable of inheriting traits from foreign DNA and the ones that are able to do so are referred to as competent (Lorenz & Wackernagel). In nature this happens by a cell absorbing free extracellular DNA from the environment, transferring the genes from the DNA to the cell. The plasmids that are taken up can be readily expressed in the cell and integrate with the cell's current DNA (Lorenz & Wackernagel 1994). Microbial DNA is found in extracellular fluid in almost every environment on earth and this process is very common in the natural world, contributing to more diversity in the gene pool (Lorenz & Wackernagel 1994). In this experiment we will be subjecting Escherichia coli, a competent bacteria, to the pGLO
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We will be using the heat shock method in this experiment, in which we will subject the cells to a sudden change in temperature, from cold to hot, in turn making the cell membrane more permeable and accepting of the foreign material (Chen & Dubnau 2004). Subjecting the bacteria to CaCl2 before the temperature change helps to facilitate the transfer of DNA into the cytoplasm for reasons that are still unknown. It is speculated that the DNA enters the cell's cytoplasm by entering through the membrane with the help of the Ca 2+ cation (Rahimzadeh et al. 2016). If successful in genetically transforming E. coli with the pGLO plasmid we should see a new found resistance to ampicllin and fluorescent properties under UV light in conjunction with the presence of arabinose sugar due to the inheritance of the green fluorescent protein (GFP) (Weedman 2016) …show more content…
coli containing the pGLO plasmid glows in the presence of arabinose under UV light
Discussion:
Any bacteria that is competent should be able to inherit traits from foreign DNA (Chen & Dubnau 2004). In this experiment we expected competent E. coli bacteria to inherit traits from the pGLO plasmid after being subjected to the heat shock method (Weedman 2016). If the transformation was successful the new DNA would give the E. coli resistance to ampicillin and the green fluorescent protein which would cause the bacteria to fluoresce under UV light in the presence of arabinose (Weedman 2016). During our experiment we were unable to definitively say if the genetic transformation had actually taken place due to a high level of experimental error in the classroom. None of our growth plates had any bacterial colonization, including the control, which indicated that something other than experimental parameters interfered with the well being of the colonies. Due to this there was no distinguishable difference between the E. coli +pGLO and the E. coli
We will be using the heat shock method in this experiment, in which we will subject the cells to a sudden change in temperature, from cold to hot, in turn making the cell membrane more permeable and accepting of the foreign material (Chen & Dubnau 2004). Subjecting the bacteria to CaCl2 before the temperature change helps to facilitate the transfer of DNA into the cytoplasm for reasons that are still unknown. It is speculated that the DNA enters the cell's cytoplasm by entering through the membrane with the help of the Ca 2+ cation (Rahimzadeh et al. 2016). If successful in genetically transforming E. coli with the pGLO plasmid we should see a new found resistance to ampicllin and fluorescent properties under UV light in conjunction with the presence of arabinose sugar due to the inheritance of the green fluorescent protein (GFP) (Weedman 2016) …show more content…
coli containing the pGLO plasmid glows in the presence of arabinose under UV light
Discussion:
Any bacteria that is competent should be able to inherit traits from foreign DNA (Chen & Dubnau 2004). In this experiment we expected competent E. coli bacteria to inherit traits from the pGLO plasmid after being subjected to the heat shock method (Weedman 2016). If the transformation was successful the new DNA would give the E. coli resistance to ampicillin and the green fluorescent protein which would cause the bacteria to fluoresce under UV light in the presence of arabinose (Weedman 2016). During our experiment we were unable to definitively say if the genetic transformation had actually taken place due to a high level of experimental error in the classroom. None of our growth plates had any bacterial colonization, including the control, which indicated that something other than experimental parameters interfered with the well being of the colonies. Due to this there was no distinguishable difference between the E. coli +pGLO and the E. coli