Introduction to the Workshop
URLs Provided by Attendees

Abstracts

Mapping

Informatics

Sequencing

Instrumentation

Ethical, Legal, and Social Issues

Infrastructure

The electronic form of this document may be cited in the following style:
Human Genome Program, U.S. Department of Energy, DOE Human Genome Program Contractor-Grantee Workshop IV, 1994.

Abstracts scanned from text submitted for November 1994 DOE Human Genome Program Contractor-Grantee Workshop. Inaccuracies have not been corrected.

Generation of Transgenic Mice Carrying Centromeric Sequences

Ana V. Perez-Castro, Julie Wilson, Michael Altherr and Robert K. Moyzis
Life Sciences Division, LS-2, MS-M880, and Center for Human Genome Studies; Los Alamos National Laboratory, Los Alamos NM 87545.

The centromere of mammalian and other complex eukaryotic chromosomes is characterized by the presence of varying amounts of nontranscribed repetitive DNA sequence.

The human satellite repeat (GGAAT)n is similar to the central region of the yeast centromere sequence CDE III. We have done PCR experiments which show that this repeat is also present in mice, flies and sea urchin, suggesting evolutionary conservation. It has been proposed that this specific repeat (GGAAT)n, could be a component of the functional centromere (1).

In order to test this hypothesis in vivo, and to see any possible effects that this sequence might have on development and gene expression, we decided to introduce a 158 bp DNA fragment containing the human (GGAAT)n repeat in the mouse genome, using pronuclear injection.

We have generated 7 independent transgenic mouse lines carrying different number of copies of the (GGAAT)n repeat. The mice are apparently normal and we are currently mating them to see if their breeding capability has been affected, and if the second generation shows any abnormality. We will present our results and discuss the significance of our findings. We are also in the process of injecting other interesting centromeric repeats and evaluating their effects in the development of fertilized mouse oocytes.

(1) Grady, L., D. et al. (1992) Highly conserved repetitive DNA sequences are present at human centromeres. Proc. Natl. Acad. Sci. USA, 89, 1695 - 1699.