chapter 20 dna technology & genomics. slide 2 of 14 biotechnology terms biotechnology process of...
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Chapter 20
DNA Technology & Genomics
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Biotechnology Terms
Biotechnology Process of manipulating organisms or their components to
make useful products Genetic engineering + tissue/cell culturing technologies
Genetic Engineering Manipulation of genes and genome Insulin & GMO (Genetically Modified Organism)
Recombinant DNA Artificially created DNA Typically, DNA is integrated from another species
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Biotechnology Terms
Gene Cloning Laboratory production of multiple copies of DNA segment Therapeutic cloning – embryonic stem cells
Spinal cord injuries Reproductive (organismal) cloning – Dolly the sheep
Restriction Enzymes Enzymes that cut DNA at specific locations Usually, derived from bacteria Cut sites of DNA = restriction fragments Sticky ends – restriction fragments usually have one end
longer than the other
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Cloning is fun!!
5 steps1. Identify & isolate the gene of interest
Involves finding a cloning vector – plasmid or organism used to carry the DNA sequence to be cloned
2. Restriction enzymes cut gene of interest from original site & cut vector’s DNA This ensures matching sticky ends on gene of interest &
vector DNA
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Cloning is fun!! (Page 2)
5 steps3. Form the recombinant plasmid
Recombinant plasmid – plasmid + DNA fragments Sealed together using DNA Ligase Restriction enzymes cut gene of interest from original site & cut
vector’s DNA This ensures matching sticky ends on gene of interest &
vector DNA
4. Get the vector (w/gene of interest) into a host cell Usually involves bacterial transformation
5. Select for cells that have been transformed Identify cells with gene of interest (antibiotic resistance)
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PCR Polymerase Chain Reaction Thermal cycling Amplification of DNA
3 Steps Denaturation (Heating)
Annealing (Cooling)Primer formation
Extension DNA polymerase adds nucleotides at 3’ end
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Biotechnology Terms (More)
Genomic library Thousands of recombinant plasmid clones Each has a piece of the original genome
cDNA Library Complementary DNA Reverse transcriptase makes cDNA from mRNA
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