Type II Estrogen Receptor in the Development and Progression of Breast Cancer.
Final rept. 1 Oct 94-30 Sep 96,
MARYLAND UNIV BALTIMORE
Pagination or Media Count:
We have purified a low affinity from of estrogen receptor type II estrogen binding site, EBS which is distinct in its steroid and DNA binding properties from the type I estrogen receptor. Purified type II EBS was utilized to generate polyclonal antibody against this protein in mice. The antibody was able to detect the type II EBS in Western Blot. In order to identify the gene encoding the type II EBS, a cDNA library was created in lambdagt11 using cDNA prepared from pregnant rat uterine tissue. The library was subsequently screened using the type II EBS antibody prepared in mouse. Although two putative clones were initially isolated, they were determined to be false positives following subsequent purification and re-screening. These results seen to indicate that the antibody was of insufficient titer for use in screening the uterine cDNA library. It is possible that rate type II EBS displays low level immunogenicity in mice. Future goals for this project include induction of expression of type II EBS in mice using estrogen implants, followed by the development of a higher yield purification procedure, which should aid in the identification of the type II EBS gene.
- Genetic Engineering and Molecular Biology
- Medicine and Medical Research