Presentation is loading. Please wait.

Presentation is loading. Please wait.

DrM4ScienceGrinds 1. Isolation 2. Cutting 3. Ligation and Insertion 4. Transformation 5. Cloning & Expression.

Similar presentations


Presentation on theme: "DrM4ScienceGrinds 1. Isolation 2. Cutting 3. Ligation and Insertion 4. Transformation 5. Cloning & Expression."— Presentation transcript:

1 DrM4ScienceGrinds 1. Isolation 2. Cutting 3. Ligation and Insertion 4. Transformation 5. Cloning & Expression

2 DrM4ScienceGrinds Donor DNA Genetic probe (a) Isolation of a specific gene from donor e.g. human Cells broken open Genetic probe added Reveals position of the gene of interest Position of gene of interest

3 DrM4ScienceGrinds  (b) Isolation of plasmid from a bacterial cell Bacterial cell Plasmid

4 DrM4ScienceGrinds  Human DNA and plasmid DNA are cut open using the same restriction enzymes From Leaving Cert Biology

5 DrM4ScienceGrinds  DNA is cut into fragments using restriction enzymes.  One RE cuts at GAATTC  DNA from two different organisms cut with GAATTC RE, cut ends from both sources are complimentary but bind weakly to each other

6 DrM4ScienceGrinds  The target gene is placed in the DNA of the plasmid/cloning vector and joins on to it  When cut plasmids are mixed with cut human DNA, different combinations result.  DNA ligase is used to form strong bonds within the recombinant DNA From Leaving Cert Biology

7 DrM4ScienceGrinds  Gets foreign DNA to join to DNA in cloning vector  Can only work if both sources of DNA have been treated with the same restriction enzymes as cut ends will be complementary to each other  Sections of human DNA can be combined with plasmid DNA which has been cut open.  DNA Ligase forms recombinant DNA From Leaving Cert Biology

8 DrM4ScienceGrinds  Uptake of recombinant DNA into cell Vast majority of cells Some cells Some cells  Special techniques will identify the small number of bacteria with the target gene From Leaving Cert Biology

9 DrM4ScienceGrinds Cloning: Identical copies of the bacteria with the target gene are produced Expression: Getting the organism with the recombinant DNA to produce the desired protein


Download ppt "DrM4ScienceGrinds 1. Isolation 2. Cutting 3. Ligation and Insertion 4. Transformation 5. Cloning & Expression."

Similar presentations


Ads by Google