International Mammalian Genome Society

The 15th International Mouse Genome Conference (2001)


POSTER 180 - THE CLONING OF A PROMOTER TRAP INSERTION SITE USING RACE AND INVERSE PCR IN THE MOUSE

Dr. Andrew Thliveris
University of Wisconsin
McArdle Laboratory for Cancer Research
1400 University Avenue
Madison, WI
53706
USA.

Co-Authors: Dove WF
Institutions: University of Wisconsin

We describe the cloning of a retroviral derived insertion site of the lacZ transgenic line Gt(ROSA)11S or (R11).   The  R11 line was generated using a retroviral promoter trap vector in ES cells. Mice from the R11 line have been shown to express lacZ ubiquitously in the embryo and hematopoietic system.  However, R11 is particularly interesting in its expression pattern in the intestinal epithelium,  expressing only in the lower proliferative zone of the crypt. There is no detectable lacZ activity in the upper region of intestinal crypts or in the surface epithelium; perhaps R11 marks at least the intestinal stem cell population. We have cloned the R11 insertion site with an inverse PCR protocol using unique retroviral vector sequences as primers. The sequence obtained from inverse PCR products identified a single mouse BAC clone (RP23-169K7) mapping to chromosome 15.  Genetic mapping of the R11 lacZ character independently localized to chromosome 15. The cDNA sequence of R11 was then identified using RACE PCR identifying a 5 prime - cDNA sequence with homology to Heterochromatin protein 1 alpha (HP-1a). This cDNA sequence also maps to the RP23-169K7 BAC clone. We intend to explore the use of markers such as R11 in dissecting the cellular pathways of intestinal neoplasia in the mouse and comparing those pathways in the human.


Abstracts * Officers * Bylaws * Application Form * Meeting Calendar * Contact Information * Home * Resources * News and Views * Membership

Base url http://imgs.org
Last modified: Saturday, November 3, 2012