Chromosome, Microdissection and Microcloning
45
clones that are within a million base pairs
of the disease locus have been isolated,
other molecular biological techniques may
be used to isolate the gene of interest.
As an example, chromosome microdissec-
tion was used to isolate a novel oncogene
(eIF-5A2 initiation factor) from human
ovarian cancer. In this disease ampli±-
cation of 3q25–q26 constitutes one of
the most frequent chromosomal alter-
ations. By hybrid selection using 3q26
band-speci±c DNA, transcribed sequences
(among which eIF-5A2) were isolated from
a primary ovarian cancer containing high-
copy number 3q26.
Table 1
lists
a
number
of
disease-
related genes that were isolated by using
microdissection and cloning methods.
Similar chromosome band–speci±c ex-
pression fragments were obtained from
probe pools generated by microdissection
of murine, bovine, swine, equine, and rice
chromosomes. The pool DNA was used
as probes to screen cDNA libraries of the
respective species.
6.4
Study of Chromosome Abnormalities in
Cancer Cells
Many nonrandom chromosome abnor-
malities are associated with speci±c cancer
cell types. Microdissection may help ad-
vance our knowledge about the structure
of such aberrant chromosomes and their
behavior during mitosis, as well as de-
lineate cytogenetic aberrations that cannot
be identi±ed by conventional chromosome
analysis. Translocation sites may be micro-
dissected directly and cloned to expedite
the isolation of the causative chimeric
(fusion) gene. Also, regions correspond-
ing to a deletion site may be dissected
from the normal chromosome. Clones
may be used as probes in
in situ
hybridiza-
tion experiments to delineate whether the
Tab. 1
Recombinant DNA libraries from microdissected chromosome
regions associated with inheritable disorders in human.
Disease
Chromosome region
Adenomatous polyposis coli
5q21
Beckwith–Wiedmann syndrome
11p15.5–pter
Cystic Fbrosis
7q
Down’s syndrome
21q
±ragile X syndrome
Xq27–Xq28
Langer–Giedion syndrome
8q24.1
Meningioma
22q12–q13
NeuroFbromatosis type II
22
Prader–Willi syndrome
15q11.1–12
Trichorhinophalangeal syndrome type I
8q23–qter
Wilms’ tumor/anoridia
11p13
Waardenburg syndrome type I
2q33–qter
Multiple endocrine neoplasia type 2A
10p11.2–q21.1
Small-cell lung cancer and renal cell carcinoma
3p14–p21
Angelman syndrome
16p11.2
Ovarian cancer
3q25–q26
Acute nonlymphocytic leukemia
4p
Germ cell Tumors
12p
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