• Harris, Stephen E (PI)

Project: Research project

Project Details


The hamster ductus deferns smooth tumor cell line (DDT-1-MF-2)
contains androgen receptors and is stimulated by androgens to
grow. In the presence of the androgen methyltrienolene, the
level of androgen receptor increases to a steady state
concentration of 20 fmoles/Mug DNA (2.0 mg of receptor/10-11
cells). The androgen receptor from these cells will be
purified by a combination of preparative denaturing Slab gel
electrophoresis, affinity chromatography, photoaffinity
labelling, and preparative two-dimensional polyacrylamide gel
electrophoresis. A phage Lambdagt 11 complementary DNA library
with over 250,000 independent recombinant has been prepared
from messenger RNA, isolated from androgen stimulated DDT-1
cells. This library has been shown to contain full length
inserts to other low abundance messenger RNAs. The pure
androgen receptor will be used to determine amino acid
composition and the amino acid sequence of various peptides.
Several oligonucleotide probes (20-40 nucleotides long) will be
prepared with minimal codon redundancy. These probes will be
used to screen the Lambdagt 11-cDNA library and to isolate and
sequence the cDNa for the androgen receptor mRNA. The cDNA
will then be used to isolate the genomic fragments for the
androgen receptor gene(s) and identify the promoter region(s).
The cDNA for this hamster androgen receptor mRNA will also be
utilized to screen a Lambdagt 11-cDNA library made to human
benign prostate hyperplasia messenger RNA. The cDNAs will then
be used to study androgen receptor gene expression in growth
and differentiation in both normal and abnormal tissues and in
cells in a variety of androgen responsive systems.
Effective start/end date7/1/866/30/89


  • National Institutes of Health


  • Medicine(all)


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