4.4 biotechnology - gene transfer (by kyung)

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Page 1: 4.4 Biotechnology - Gene transfer (by Kyung)

Gene Transfer Kyung Cho

B Block

Page 2: 4.4 Biotechnology - Gene transfer (by Kyung)

4.4.8O Outline a basic technique used for

gene transfer involving plasmids, a host cell (bacterium, yeast or other cell), restriction enzymes (endonucleases) and DNA ligase.

Page 3: 4.4 Biotechnology - Gene transfer (by Kyung)

Words to knowO Gene transfer – Gene is carried by a conjugative plasmid.

The process is essential for fertility and establishment of the bacterial donor state

O Host cell - A cell that has been introduced with DNA (or RNA), such as a bacterial cell acting as a host cell for the DNA isolated from a bacteriophage

O Restriction enzymes- An enzyme that catalyzes the cleavage of DNA at restriction sites, producing small fragments used for gene splicing in recombinant DNA technology and for chromosome mapping

O DNA ligase – DNA ligase joins the sticky ends, fixing the gene into the E.coli plasmid

O Plasmids - A linear or circular double-stranded DNA that is capable of replicating independently of the chromosomal DNA

Page 4: 4.4 Biotechnology - Gene transfer (by Kyung)

Process

1. Restriction enzymes cut out gene that is to be transferred

2. The same restriction enzyme cuts into the plasmid

3. Because it is the same restriction enzyme, the same complementary 'sticky ends' in the plasmid

Page 5: 4.4 Biotechnology - Gene transfer (by Kyung)

Process4. DNA ligase joins the sticky ends, fixing the gene into the E.coli plasmid5. The recombinant plasmid is

inserted into the host cell. 6. It now expresses the new gene.

Page 6: 4.4 Biotechnology - Gene transfer (by Kyung)

An example

Page 8: 4.4 Biotechnology - Gene transfer (by Kyung)
Page 9: 4.4 Biotechnology - Gene transfer (by Kyung)

The End