Dr. Mohamed El-AziziDepartment of Microbiology andBiotechnology
Cloning and expression of insulin gene
Biotechnology: Demystifying the Concepts p (138-168)
Compensatory Lectures
Lecture #Original dateCompensationdateSlot/LocationMicrobiology712/4/20095/4/20092
nd
/H5Microbiology819/4/200921/4/20095
th
/H5Introduction toBiotechnology412/4/200914/4/20095
th
/H5
The cloned gene can be detected by a technique known as Colony hybridization
:1-Cells from each colonies are transferred to a filter paper by pressing the paper against the surface of the agar plate.2-Treating the paper with denature solution which lyses the cellsand releases the DNA.3-Denatured DNA adheres to the filter.4-The filter is treated with a labeled probe (specific to the target gene) to allow hybridization betweenthe probe and the denatured DNA of the gene (if it exists).5-The filter is washed with fresh solution to remove the excess free probes (not bound specifically todenatured DNA).6-Emission of the labeled probe forms black spot on X-ray film.7. Comparison of the location of the probe signal on the film tothe location of the colony on the originalagar plate identifying colony containing DNA complementary to the probe (target gene)
oos o ene onng4. Detection of the Cloned Gene by Colony
Hybridization Technique
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Leture 3. Cloning and Expression of Insulin Gene in Bacteria
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