Recombinant DNA and Polymerase chain reaction

Page
2

Slide 9

How do we know which bacteria have the gene?

It is necessary to isolate the host bacteria that contain the gene that has been spliced as only want the recombinant DNA

By having a gene on the same plasmid that gives resistance to an antibiotic, the other bacteria can be removed by culturing the bacteria in a medium that contains the antibiotic.

The bacteria containing the resistance to the antibiotic will survive and the others will be killed by the antibiotic.

Slide 10

Selection of Altered Cells

Antibiotic resistance gene used to identify recombinant cells

Slide 11

http://www.sumanasinc.com/webcontent/animations/content/plasmidcloning.html

ANIMATION – RECOMBINANT DNA

Slide 12

Plasmids will not work as well in eukaryotic organisms like plants and animals

Other methods need to be used to insert the DNA

Viral vectors can be used for animal cells.

The virus can ‘inject’ their DNA into an animal host cell.

Can we use this technique on all cells?

Slide 13

Gene Gun can be used to insert genes into plant cells

http://www.hort.purdue.edu/hort/courses/HORT250/animations/Gene%20Gun%20Animation/Genegun1.html

GENE GUN

Slide 14

Biotechnologists are Problem Solvers!

Diabetics having reactions to porcine/animal insulin

Wheat crops being attacked by insects

People sick with cystic fibrosis

All these can be fixed by recombinant DNA!!!

On a Flow Chart show the steps involved in making recombinant DNA for a desired gene. From cutting of the gene to the final product (this may involve the delivery method)

Slide 15

Now we have made the gene – how do we get lots of copies??

Slide 16

E.Coli Plasmid is cut with the same restriction enzyme used to cut the insulin gene

Insulin gene is cut from a pancreatic cell DNA using a specific restriction enzyme

insulin - Bacterial cells when supplied with required polypeptides or proteins, the colonies will produce insulin

E.g Vaccines- The plamids are isolated from the e.coli cells, the genes are then amplifyed via PCR and used to create inactivated viruses for vaccines

Slide 17

Useful Properties of DNA