recombinant dna and genetic engineering chapter 12
TRANSCRIPT
Recombinant DNA and Genetic Engineering
Recombinant DNA and Genetic Engineering
Chapter 12Chapter 12
Goals for this chapterGoals for this chapter
Have a general understanding of Genetic Engineering and Biotechnology
Be able to form an opinion that is based on fact or emotion
And recognize the difference
Have a general understanding of Genetic Engineering and Biotechnology
Be able to form an opinion that is based on fact or emotion
And recognize the difference
Focus our attention onFocus our attention on
Genetic Improvement of Crop Plants
Other possible topics Genetic improvement of animals Genetic improvement of Humans Stem cell therapy Forensics Historical research And on and on
Genetic Improvement of Crop Plants
Other possible topics Genetic improvement of animals Genetic improvement of Humans Stem cell therapy Forensics Historical research And on and on
What is BiotechnologyWhat is Biotechnology
Definitions
Biotechnology Genetic Engineering Recombinant DNA Transgenic or GMO
Definitions
Biotechnology Genetic Engineering Recombinant DNA Transgenic or GMO
BiotechnologyBiotechnology
Genetic Improvement of plants and animals This would include the activities of Plant
Breeders
And plant breeders are really nice people
Genetic Improvement of plants and animals This would include the activities of Plant
Breeders
And plant breeders are really nice people
Genetic ChangesGenetic 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
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
Traditional plant breedingTraditional plant breeding
Select parents Cross and generate variable offspring Select the desired types Test - Test - Test Multiply Release 7 to 15 years
Select parents Cross and generate variable offspring Select the desired types Test - Test - Test Multiply Release 7 to 15 years
Norman BorlaugNorman Borlaug
Nobel Peace Prize 1970 The Green Revolution
http://nobelprize.org/peace/laureates/1970/borlaug-lecture.html
Nobel Peace Prize 1970 The Green Revolution
http://nobelprize.org/peace/laureates/1970/borlaug-lecture.html
Barbara MclintockBarbara Mclintock
Transposons 1983 Nobel Prize
in Medicine
Transposons 1983 Nobel Prize
in Medicine
Genetic EngineeringGenetic Engineering
Modern Biotechnology - which uses the knowledge of DNA to manipulate the genetic makeup of an organism
Recombinant DNA - take a gene from one organism and place it into another organism
Transgenic or GMO - an organism that contains DNA from another organism
Modern Biotechnology - which uses the knowledge of DNA to manipulate the genetic makeup of an organism
Recombinant DNA - take a gene from one organism and place it into another organism
Transgenic or GMO - an organism that contains DNA from another organism
Would You Eat a Genetically EngineeredFood?
Would You Eat a Genetically EngineeredFood?
Most GMO’s are PlantsMost GMO’s are Plants
34% of Corn 71% of Cotton 75% of soybeans
34% of Corn 71% of Cotton 75% of soybeans
Silk is SoySilk is Soy
Why no GMOWhy no GMO
Until recently the terms Genetically Modified Organism (GMO), GMO-Free and Non-GMO were used to help identify foods that contained genetically altered ingredients.
These terms are no longer recognized by the Food and Drug Administration (FDA) and therefore cannot be used on food packaging.
Until recently the terms Genetically Modified Organism (GMO), GMO-Free and Non-GMO were used to help identify foods that contained genetically altered ingredients.
These terms are no longer recognized by the Food and Drug Administration (FDA) and therefore cannot be used on food packaging.
What about Tostito’sWhat about Tostito’s
Or Pepsi Or Coke Or CapN Crunch???
Or Pepsi Or Coke Or CapN Crunch???
How to Manipulate DNAin the Lab
How to Manipulate DNAin the Lab
Examples of TransformationExamples of Transformation Natural Systems
Bacteria Viruses
Natural Systems Bacteria Viruses
Bacterium
bacterialchromosome
plasmid
Transformation with DNA fragment
bacterialchromosome
DNAfragments
Virus enters host cell.2
virus
viral DNA
host cell
host cell DNA
“hybrid virus” viral proteins
viral DNA
Virus attaches tosusceptible host cell.1
Virus releases its DNA intohost cell; some viral DNA (red)may be incorporated into thehost cell’s DNA (blue).
3
Viral genes encode synthesisOf viral proteins and viral geneReplication. Some host cell DNAMay attach to replicated viralDNA (red/blue).
4
New viruses assemble; hostcell DNA is carried by “hybridviruses.”
5
Host cell bursts, releasingnewly assembled viruses. when “hybrid viruses” infect asecond cell, they may transfergenes from the first cell to thesecond cell.
6
GE Tool BoxGE Tool Box
Restiction Enzymes Cloning Vectors cDNA Cloning Reverse
Transcriptase PCR
Restiction Enzymes Cloning Vectors cDNA Cloning Reverse
Transcriptase PCR
Gene Library (Isolation)
Transformation of Plants
Gene Library (Isolation)
Transformation of Plants
Based on the Central Dogma and the fact that in Based on the Central Dogma and the fact that in virtually organisms the CODE is perfectly conserved virtually organisms the CODE is perfectly conserved almostalmost
Amplifying DNAAmplifying DNA
Fragments can be inserted into fast-growing microorganisms
Polymerase chain reaction (PCR)
Fragments can be inserted into fast-growing microorganisms
Polymerase chain reaction (PCR)
Polymerase Chain ReactionPolymerase Chain Reaction Sequence to be copied is heated
Primers are added and bind to ends of single strands
DNA polymerase uses free nucleotides to create complementary strands
Doubles number of copies of DNA
Sequence to be copied is heated
Primers are added and bind to ends of single strands
DNA polymerase uses free nucleotides to create complementary strands
Doubles number of copies of DNA
Polymerase Chain Reaction
Polymerase Chain Reaction
Double-stranded DNA to copy
DNA heated to 90°– 94°C
Primers added to base-pair with ends
Mixture cooled; base-pairing of primers and ends of DNA strands
DNA polymerasesassemble new DNA strands
Fig. 16-6, p. 256
Stepped Art
Polymerase Chain Reaction
Polymerase Chain Reaction
Stepped Art
Mixture heated again; makes all DNA fragments unwind
Mixture cooled; base-pairing between primers and ends of single DNA strands
DNA polymerase action again doubles number of identical DNA fragments
Fig. 16-6, p. 256
Fig. 16-7, p.247
DNA FingerprintingDNA Fingerprinting
Guilty or InnocentGuilty or Innocent
8 side-by-side (tandem) repeatsof the same 4-nucleotide sequence,
Nylon paper with DNA is bathed in a solution of labeled DNA probes (red) that are complementary to specific DNA segments in the original DNA sample.
Complementary DNA segments are labeled by probes(red bands).
nylon paper
solution of DNA probes (red)
gel
gel
power supply
DNA bands(not yet visible)
� �
wells
pipetter
nylonpaper
DNA samples are pipetted into wells(shallow slots) in the gel. Electrical currentis sent through the gel (negative at endwith wells, positive at opposite end).
Electrical current moves DNAsegments through the gel. Smallerpieces of DNA move farther toward thepositive electrode.
Gel is placed on special nylonpaper. Electrical current drivesDNA out of gel onto nylon.� �
gel
power supply
� �
wells
pipetter
DNA samples are pipetted into wells(shallow slots) in the gel. Electrical currentis sent through the gel (negative at endwith wells, positive at opposite end).
� �
DNA bands(not yet visible)
� �
Electrical current moves DNAsegments through the gel. Smallerpieces of DNA move farther toward thepositive electrode.
� �
gel� �
nylonpaper
Gel is placed on special nylonpaper. Electrical current drivesDNA out of gel onto nylon.
� �
Nylon paper with DNA is bathed in a solution of labeled DNA probes (red) that are complementary to specific DNA segments in the original DNA sample.
nylon paper
solution of DNA probes (red)� �
� �
Complementary DNA segments are labeled by probes(red bands).
� �
� �
STR name
Penta D
CSF
D16
D7
D13
D5
D16: an STR on chromosome 16
DNA samples from13 different people
Num
ber
of r
epea
ts
STR name
Penta D
CSF
D16
D7
D13
D5
Num
ber
of r
epea
ts
D16: an STR on chromosome 16
DNA samples from13 different people
Num
ber
of r
epea
ts
Genetic EngineeringTransformation of PlantsGenetic EngineeringTransformation of Plants Genes are isolated, modified, and
inserted into an organism Made possible by recombinant
technology
Cut DNA up and recombine pieces
Amplify modified pieces
Genes are isolated, modified, and inserted into an organism
Made possible by recombinant technology
Cut DNA up and recombine pieces
Amplify modified pieces
ProcessProcess
Board Diagram
Corn Transformation with RR gene
From a bacteria or petunia
Hypothetical Situation
Corn Cotton and Alfalfa
Board Diagram
Corn Transformation with RR gene
From a bacteria or petunia
Hypothetical Situation
Corn Cotton and Alfalfa
Roughly 400 million people in the world today are at risk of Vitamin A deficiency, which already affects 100-200 million children. Vitamin A deficiency causes various health problems, including blindness. Because rice is an important crop, eaten by almost half of the people in the world, the Rockefeller Foundation and the European Union funded research into varieties that might offer global health benefits.
It may now be possible, thanks to agricultural biotechnology, to make rice and other crops into additional sources of Pro-Vitamin A. With Monsanto's help, the developers of "Golden Rice" and mustard with more Pro-Vitamin A should one day be able to deliver their gift of better nutrition to the developing nations of the world through staple crops readily available to poor and vulnerable populations
Imagine sharing science to help others develop crops that could help reduce Vitamin A deficiency, a leading cause of blindness and infection among the young.
Imagine innovative agriculture that creates incredible things.
DiscussionDiscussion
Debate the Merits or Dangers of Golden Rice.
Group One - Make your case in front of Congress to obtain money to distribute this rice (as rice seed) to the farmers of Southeast Asia and Africa
Group Two - Argue against this.
Debate the Merits or Dangers of Golden Rice.
Group One - Make your case in front of Congress to obtain money to distribute this rice (as rice seed) to the farmers of Southeast Asia and Africa
Group Two - Argue against this.
You be the judgeYou be the judge
With your pocket book Or your advocacy
With your pocket book Or your advocacy
Ethical IssuesEthical Issues
Who decides what should be “corrected”
through genetic engineering?
Should animals be modified to provide
organs for human transplants?
Should humans be cloned?
Who decides what should be “corrected”
through genetic engineering?
Should animals be modified to provide
organs for human transplants?
Should humans be cloned?