Protein specificity of the hyperactive mutant mmPing20
School Name
Governor's School for Science and Mathematics
Grade Level
12th Grade
Presentation Topic
Microbiology
Presentation Type
Mentored
Oral Presentation Award
1st Place
Abstract
Transposable elements are DNA capable of movement within the genome. The rate at which the transposon moves (transposition rate) can be determined. mPing, a transposon found in rice, and a mutant of mPing, mmPing20, with 7 interior base pair differences both require proteins to enable movement. Previous research shows that mmPing20 transposes at roughly 1.5x the rate of mPing when a protein construct combining parts of Ping and Pong proteins was used. By using one protein type at a time (either Ping or Pong), we can determine if mmPing20’s hyperactivity is specific to Ping or Pong proteins and how they react to the base pair changes. To test this we utilized yeast transposition assays with yeast transformed with a transposon (either mPing or mmPing20) interrupting the ADE2 gene producing adenine, needed by yeast to grow. The yeast colonies growing on the transposition assay plates were counted and the transposition rate was calculated. The experiments showed that when Ping proteins were used, mmPing20 transposed at 2.76x that of mPing but when Pong proteins were used, mmPing20 transposed at 1.80x that of mPing. ORF1 proteins are thought to bind to the interior of transposons so since mmPing20’s base pair differences were in its interior, the binding of the Ping ORF1 must be affected. Further, the transposition rates of mPing and mmPing20 when Pong proteins were used indicated that Pong ORF1’s binding behavior was less affected by the 7bp differences than Ping since the rates were not as different.
Recommended Citation
Bombard, David, "Protein specificity of the hyperactive mutant mmPing20" (2018). South Carolina Junior Academy of Science. 65.
https://scholarexchange.furman.edu/scjas/2018/all/65
Location
Neville 221
Start Date
4-14-2018 8:45 AM
Presentation Format
Oral and Written
Protein specificity of the hyperactive mutant mmPing20
Neville 221
Transposable elements are DNA capable of movement within the genome. The rate at which the transposon moves (transposition rate) can be determined. mPing, a transposon found in rice, and a mutant of mPing, mmPing20, with 7 interior base pair differences both require proteins to enable movement. Previous research shows that mmPing20 transposes at roughly 1.5x the rate of mPing when a protein construct combining parts of Ping and Pong proteins was used. By using one protein type at a time (either Ping or Pong), we can determine if mmPing20’s hyperactivity is specific to Ping or Pong proteins and how they react to the base pair changes. To test this we utilized yeast transposition assays with yeast transformed with a transposon (either mPing or mmPing20) interrupting the ADE2 gene producing adenine, needed by yeast to grow. The yeast colonies growing on the transposition assay plates were counted and the transposition rate was calculated. The experiments showed that when Ping proteins were used, mmPing20 transposed at 2.76x that of mPing but when Pong proteins were used, mmPing20 transposed at 1.80x that of mPing. ORF1 proteins are thought to bind to the interior of transposons so since mmPing20’s base pair differences were in its interior, the binding of the Ping ORF1 must be affected. Further, the transposition rates of mPing and mmPing20 when Pong proteins were used indicated that Pong ORF1’s binding behavior was less affected by the 7bp differences than Ping since the rates were not as different.
