Cell Division in Bacteria
- Submitted by: trevorthompson03
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- Category: Science
- Date Submitted: 11/29/2010 12:16 PM
- Pages: 6
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The overall purpose of this study was to determine and understand cell divisions in bacteria, specifically the septation process and the proteins involved. The researchers experimented to discover which proteins were involved in cell division and when. They focused on fts genes and their mutations to better understand the protein complexes involved in cell division of bacteria. Through confirmation of the localization of the proteins they support their idea that a large protein complex functions in septation. To support their hypothesis, they clone the fts gene and its mutations to view its localization activity
The researchers want to understand what proteins were involved, and when they were involved, in cell division, they used PCR. PCR is a common technique in microbiology used to amplify a specific piece of DNA to make thousands to millions of copies of a particular DNA sequence. PCR is used to amplify ftsK fragments in this experiment, and therefore specific primers are necessary. They made specific primers FtsK1, FtsK3, FtsK4, and FtsK7; each primer keeps the same N-terminus intact, Ftsk1, and then made a different length, FtsK3 being the longest and FtsK7 being the shortest, in order to see what part of the FtsK protein targets . The N-terminus is the start of a protein. They created different lengths, having the same fusion junction with GFP, but varying on the 3’ end, in order to the amount of amino acids needed to target GFP to septum. The primer for FtsK1 included a Sac 1 site so that when inserted into the plasmid it would be inserted right in front of the promoter lacZ. The other three included an XbaI site at the desired end so that the GFP, also cut with the XbaI, would connect. Gfp is a gene that produces a protein, GFP, that when turned on will glow. Therefore, by tagging the FtsK, one can see what it is doing, and when it is doing it. In this case, GFP is turned on by the lac promoter. Once primers were created, in order to amplify the...
The researchers want to understand what proteins were involved, and when they were involved, in cell division, they used PCR. PCR is a common technique in microbiology used to amplify a specific piece of DNA to make thousands to millions of copies of a particular DNA sequence. PCR is used to amplify ftsK fragments in this experiment, and therefore specific primers are necessary. They made specific primers FtsK1, FtsK3, FtsK4, and FtsK7; each primer keeps the same N-terminus intact, Ftsk1, and then made a different length, FtsK3 being the longest and FtsK7 being the shortest, in order to see what part of the FtsK protein targets . The N-terminus is the start of a protein. They created different lengths, having the same fusion junction with GFP, but varying on the 3’ end, in order to the amount of amino acids needed to target GFP to septum. The primer for FtsK1 included a Sac 1 site so that when inserted into the plasmid it would be inserted right in front of the promoter lacZ. The other three included an XbaI site at the desired end so that the GFP, also cut with the XbaI, would connect. Gfp is a gene that produces a protein, GFP, that when turned on will glow. Therefore, by tagging the FtsK, one can see what it is doing, and when it is doing it. In this case, GFP is turned on by the lac promoter. Once primers were created, in order to amplify the...