Event Title

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.

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Apr 21st, 10:00 AM Apr 21st, 11:30 AM

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.
http://cornerstone.lib.mnsu.edu/urs/2014/poster_session_A/19