The Cystic Fibrosis Gene

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when searching
for the CF gene. To find the gene responsible for CF, the principle of
"reverse genetics" was applied. Scientists accomplished this by linking the
disease to a specific chromosome. After this linkage, they isolated the
gene of interest on the chromosome and then tested its product.
Before the disease could be linked to a specific chromosome, a
marker needed to be found that would always travel with the disease. This
marker is known as a Restriction Fragment Length Polymorphism or RFLP for
short. RFLP's are varying base sequences of DNA in different individuals
which are known to travel with genetic disorders.3 The RFLP for cystic
fibrosis was discovered through the techniques of Somatic Cell
Hybridization and through Southern Blot Electrophoresis (gel separation of
DNA). By using these techniques, three RFLP's were discovered for CF; Doc
RI, J3.11, and Met. Utilizing in situhybridization, scientists discovered
the CF gene to be located on the long ...

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contained the CF gene. This could be
accomplished because Northern Blot electrophoresis utilizes RNA instead of
DNA. The RNA of cell types affected with CF, along with the RNA of
unaffected cell types were placed on a gel. Probe number two bound t o the
RNA of affected cell types in the pancreas, colon, and nose, but did not
bind to the RNA from non-affected cell types like those of the brain and
heart. Probe number one did not bind exclusively to cell types from
CF affected areas like probe number two did. From this evidence, it
was determined that probe number two contained the CF gene.
While isolating the CF gene and screening the genetic library made
from mRNA (cDNA ...

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