Research Project
2200048
The overall goal of the proposed study is to evaluate the use of embryonic biopsy and polymerase chain reaction (PCR) amplification to identify microinjected pronuclear embryos will be cultured to the 8-cell stage and a small sample of embryonic cells will be removed with a biopsy pipet and lysed to provide a small sample of embryonic cells will be removed with a biopsy pipet and lysed to provide a small sample of DNA for analysis. This method of genomic analysis will maintain embryo viability and is intended to restrict embryo transfer to the small subset of microinjected embryos bearing integrated transgenes. PCR results obtained from lysates of the biopsy samples will be validated by specific restriction enzyme digests designed to discriminate between integrated and non-integrated transgenes and by analysis of offspring. These techniques will be developed and evaluated in Phase I using a mouse model system. The methods resulting in the Phase I study will be utilized in Phase II of the project, in which a herd of transgenic dairy goats, expressing mammary-specific transgenes, will be developed for use as pharmaceutical bioreactors.
