16 Recombinant DNA - Recombinant DNA and Genetic...

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Recombinant DNA and Genetic Engineering Chapter 16
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Familial Hypercholesterolemia Gene encodes protein that serves as cell’s LDL receptor Two normal alleles for the gene keep blood level of LDLs low Two mutated alleles lead to abnormally high cholesterol levels & heart disease
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Example of Gene Therapy Woman with familial hypercholesterolemia Part of her liver was removed Virus used to insert normal gene for LDL receptor into cultured liver cells Modified liver cells placed back in patient
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Results of Gene Therapy Modified cells alive in woman’s liver Blood levels of LDLs down 20 percent No evidence of atherosclerosis Cholesterol levels remain high Remains to be seen whether procedure will prolong her life
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Genetic Changes Humans have been changing the genetics of other species for thousands of years Artificial selection of plants and animals Natural processes also at work Mutation, crossing over
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Genetic Engineering Genes are isolated, modified, and inserted into an organism Made possible by recombinant technology Cut DNA up and recombine pieces Amplify modified pieces
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Discovery of Restriction Enzymes Hamilton Smith was studying how Haemophilus influenzae defend themselves from bacteriophage attack Discovered bacteria have an enzyme that chops up viral DNA
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Specificity of Cuts Restriction enzymes cut DNA at a specific sequence Number of cuts made in DNA will depend on number of times the “target” sequence occurs
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Making Recombinant DNA 5’ 3’ G C T T A A A A T T C G G A A T T C C T T A A G 3’ 5’ one DNA fragment another DNA fragment 3’ 5’ In-text figure Page 254
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Making Recombinant DNA nick 5’ 3’ 3’ 5’ G A A T T C C T T A A G nick G A A T T C C T T A A G DNA ligase action In-text figure Page 254
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Using Plasmids Plasmid is small circle of bacterial DNA Foreign DNA can be inserted into plasmid Forms recombinant plasmids Plasmid is a cloning vector Can deliver DNA into another cell
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Using Plasmids DNA fragments + enzymes recombinant plasmids host cells containing recombinant plasmids Figure 16.4 Page 255
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Making cDNA mRNA transcript mRNA–cDNA hybrid single-stranded cDNA double-stranded cDNA Figure 16.5 Page 255
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Amplifying DNA Fragments can be inserted into fast-growing microorganisms Polymerase chain reaction (PCR)
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This note was uploaded on 05/19/2011 for the course BIO 20 taught by Professor Churva during the Spring '11 term at Mapúa Institute of Technology.

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16 Recombinant DNA - Recombinant DNA and Genetic...

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