Genetic Egineering Isolation Cutting Ligation and Insertion Transformation Expression

Isolation of 1. a specific gene from donor e. g Isolation of 1. a specific gene from donor e.g. gene for insulin from human 2. plasmid from a bacterial cell

Cutting Restriction enzymes act as molecular scissors and cut DNA at specific sites called restriction sites Leaving sticky ends (Complimentary unmatched base sequences Restriction site Restriction ezymes

DNA Ligase http://www.slic2.wsu.edu:82/hurlbert/micro101/pages/Chap10.html#Sticky_ended_cut Ligation –rejoining cut fragments of DNA and forming artificial recombinant molecules

Ligation and Insertion

Recombinant DNA introduced into bacterial cell 4. Transformation Recombinant DNA introduced into bacterial cell Know this term Bacterial chromosome Bacterial cell Recombinant DNA

5. Expression Bacterial cell reproduces by Binary Fisson Bacterial cell produces the polypeptide/protein Coded for by the donor DNA e.g. insulin Universality of genetic code Plasmid will produce the polypeptide coded for by the donor DNA

Now have a transgenic organism A genetically modified organsim CONCERNS Effects on health “escape” of the transferred gene esp in plants.

Homework Questions 1. What are the 5 stages in genetic engineering? 2. What do restriction enzymes do? 3. What does DNA ligase do? 4. What is the technical term for the plasmid with DNA from two species? 5. Transformation means that you put recombinant DNA where? 6. Making many bacteria from one is called ? 7. Getting the cloned bacteria to make your product is called ?

DNA profiling

Cut DNA using restriction enzymes Extract DNA Cut DNA using restriction enzymes Separate fragments using gel electrophoresis Compare band patterns from different sources