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 arm of chromosome ...

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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 to 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 library), it was discovered that probe ...

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