Optimization of the Presence of an Actin Capping Protein Transgene in Genetically Modified Mice
Location
CSU Ballroom
Start Date
21-4-2014 10:00 AM
End Date
21-4-2014 11:30 AM
Student's Major
Biological Sciences
Student's College
Science, Engineering and Technology
Mentor's Name
Marilyn Hart
Mentor's Email Address
marilyn.hart@mnsu.edu
Mentor's Department
Biological Sciences
Mentor's College
Science, Engineering and Technology
Description
Actin Capping Protein (CP) is a heterodimer composed of an alpha and beta subunit, which binds to the barbed ends of actin filaments. Three isoforms of alpha (α1, α2, α3) and beta (β1, β2, β3) subunits have been identified. To evaluate the functions of the beta isoforms, a transgenic line of mice was produced that added additional copies of the β2 isoform of actin capping protein under the control of the myosin heavy chain promoter. The transgene is expressed in the cardiac muscle post gestationally. As such, the integrity of the strain is subject to anyone involved with the colony, including caretaking and husbandry. The colony is now 14 years old and to continue research with constancy, the transgenic mice must be genotyped. Genomic DNA was isolated from wild type and transgenic by digestion of tail clippings. The DNA was purified by phenol/chloroform extraction and precipitated by the addition of ethanol. We are using Polymerase Chain Reaction (PCR) to amplify the transgene using myosin heavy chain and beta two specific primers. We have optimized the buffer, annealing temperature and other PCR variables. The transgene is approximately 1647 base pairs. An internal fatty acid binding protein (FABPI) generated a product of 466 base pairs. The results of this project will be utilized in all subsequent research of the colony as evidence of quality control in specimens.
Optimization of the Presence of an Actin Capping Protein Transgene in Genetically Modified Mice
CSU Ballroom
Actin Capping Protein (CP) is a heterodimer composed of an alpha and beta subunit, which binds to the barbed ends of actin filaments. Three isoforms of alpha (α1, α2, α3) and beta (β1, β2, β3) subunits have been identified. To evaluate the functions of the beta isoforms, a transgenic line of mice was produced that added additional copies of the β2 isoform of actin capping protein under the control of the myosin heavy chain promoter. The transgene is expressed in the cardiac muscle post gestationally. As such, the integrity of the strain is subject to anyone involved with the colony, including caretaking and husbandry. The colony is now 14 years old and to continue research with constancy, the transgenic mice must be genotyped. Genomic DNA was isolated from wild type and transgenic by digestion of tail clippings. The DNA was purified by phenol/chloroform extraction and precipitated by the addition of ethanol. We are using Polymerase Chain Reaction (PCR) to amplify the transgene using myosin heavy chain and beta two specific primers. We have optimized the buffer, annealing temperature and other PCR variables. The transgene is approximately 1647 base pairs. An internal fatty acid binding protein (FABPI) generated a product of 466 base pairs. The results of this project will be utilized in all subsequent research of the colony as evidence of quality control in specimens.
Recommended Citation
Koonst, Tyler. "Optimization of the Presence of an Actin Capping Protein Transgene in Genetically Modified Mice." Undergraduate Research Symposium, Mankato, MN, April 21, 2014.
https://cornerstone.lib.mnsu.edu/urs/2014/poster_session_A/19
