Recombinant DNR Probe Technology in the Search For the Multiple Endocrine Neoplasia Type 2B Gene
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Authors
Drabik, Christopher E.
Issue Date
1988
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
Multiple Endocrine Neoplasia (MEN) type 2A and 2B are
related diseases showing phenotypic overlap in the kinds of
tumor involvement of each disease. Recently, by linkage to DNA
markers, Multiple Endocrine Neoplasia type 2A (MEN 2A) has been
localized to the pericentric region of chromosome 10 (Simpson
and Kidd, 1987 and Mathews 1987). Because of phenotype
overlap and consistent tumor involvement, it has been proposed
that MEN type 2A and 2B should consist of overlapping genes
causing the various syndromes of the disease (Jackson and Van
Dyke et al. 1987). No evidence to date has been given showing
that MEN 2A and 2B are linked. Isolating and characterizing
either of the two loci could lead to a further understanding of
the genetic and molecular aspects of cancer. Also, isolating
markers near either of the genes could lead to better prediction
of the diseases. Although, MEN 2A has been localized by probes
to chromosome 10, the recombination frequency between the
probes and MEN 2A is sufficiently large enough for prediction of
MEN 2A to be ruled out.
The generation of probes from a recombinant DNA library and
the use of recombinant DNA probes for genetic linkage analysis
in the search for the Men 2B gene locus was explored in this
study. R cosmid library consisting of fragments of DNR from
chromosome ten inserted into the vector pCos 2 and then cloned
into E. coli was characterized using restriction enzymes. The
recombinant cosmid clones were screened for unique sequences
by hybridization to total human genomic DNA and then screened
once more against DNR from chromosome ten only. 32 probes
specific for chromosome 10 were generated from 250 cosmids.
The previously localized chromosome ten probes D 1 OZ 1,
D10S5 and RBP3 were analyzed for linkage to the Men 2B gene
locus in 8 families affected with Men 2B. DNR from every
available individual was digested with several restriction
enzymes and the resulting fragments hybridized to D 1 OZ 1, 01 OS5
and RBP3. Families with informative polymorphism for the
restriction enzymes were used to determine whether linkage to
the probes existed. Lod scores were compiled for the Men 2B
locus verse the loci represented by the probes and feasability of
linkage to the probe loci determined. D 1 OZ 1, a probe consisting
of a sequence of DNR from the centromere of chromosome 1 0,
was found to be linked to MEN 2B. Linkage analysis of D 1 OS5 and
RBP3 vs. MEN 2B was inconclusive.
Description
42 p.
Citation
Publisher
Kalamazoo College
License
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