Office of Undergraduate Research

Gene trapping as a means to create stable transgenic zebrafish lines

The objective of this project is to generate and charcterize lines of transgenic zebrafish expressing fluorescent proteins in different cell types. Single cell zebrafish embryos are injected with DNA encoding fluorescent proteins and the resulting fish are raised to adulthood. The adult fish are spawned to generate offspring which are screened using epifluorescent microscopy for expression of the fluorescent protein. In fish expressing the fluorescent protein, the insertion site of the transgene, which encodes the fluorescent protein, is determined using polymerase chain reaction (PCR) technology. The fluorescent protein is analyzed using immunoblots (Westerns) probed with an antibody specifc for the fluorescent protein. Using these techniques we will determine in what cells the fluorescent proteins are expressed (e.g. neurons, heart, liver, eye) and when during development (e.g. in the embryo, larvsa or adult) the fluorescent protein is expressed.

This project is located at the School of Freshwater Science: 600 E. Greenfield Ave. Milwaukee, WI 53204

***Campus Shuttle Service to The School of Freshwater Science is now available!***

https://www4.uwm.edu/freshwater/

Tasks and responsibilities:

The student will be involved in every step of the process from creating the transgene to injecting the one cell embryos with DNA encoding the transgene to characterizing the resulting transgenic fish. The student will assist in fish husbandry, including feeding the fish, spawning the fish and raising the babies. The student will be responsible for keeping track of the relevant information on the fish he uses and accurate record keeping of his experiments. The student will be taught how, and be expected, to treat the fish humanely. Student will be instructed in the proper use of the microscopes used for observing the fish and will be expected to make accurate observations of the fish. In addition the student will be expected to assist in doing the day to day chores necessary for a lab to work.