topic 7 learning outcomes.docx

topic 7 learning outcomes.docx - TOPIC 7 LEARNING OUTCOMES...

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TOPIC 7 LEARNING OUTCOMES 1. DNA cloning? a. Prepare gene-sized pieces of DNA in identical copies. Use bacteria & their plasmids 2. Plasmids? a. Small circular DNA molecules that replicate separately from the bacterial chromosome 3. Gene cloning? a. Foreign DNA inserted in plasmid, recombinant plasmid inserted in bacterial cell. Bacterial cell reproduces and plasmid with foreign DNA replicates. Result= multiple copies of single gene 4. Why is gene cloning important? a. Useful for making copies of particular gene and producing a protein product 5. Restriction enzymes/endonucleases? a. Cut DNA molecules at specific DNA sequences called restriction sites. b. ‘molecular scissors’ c. Defence mechanism against attack of foreign DNA from invading viruses 6. Restriction fragments a. Many cuts made by restriction enzyme 7. Restriction enzymes 3 yields? a. 5’ overhangs (sticky) b. 3’ overhangs (sticky) c. Blunt ends 8. Why are sticky yields good? a. Staggered cut. Sticky ends can bond with complementary sticky ends of other fragments 9. Palindromic? a. Restriction recognition sites are palindromic b. The site reads the same on the reverse strand as it does on the forward strand when both are read in the same orientation 10. Cloning vector a. Original plasmid b.
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