Recombinant DNA and Genetic Engineering Chapter 12

Goals 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 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 Biotechnology  Definitions  Biotechnology  Genetic Engineering  Recombinant DNA  Transgenic or GMO  Definitions  Biotechnology  Genetic Engineering  Recombinant DNA  Transgenic or GMO

Biotechnology  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 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 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 Borlaug  Nobel Peace Prize  1970  The Green Revolution  /borlaug-lecture.html  Nobel Peace Prize  1970  The Green Revolution  /borlaug-lecture.html

Barbara Mclintock  Transposons  1983 Nobel Prize in Medicine  Transposons  1983 Nobel Prize in Medicine

Genetic 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 Engineered Food?

Most GMO’s are Plants  34% of Corn  71% of Cotton  75% of soybeans  34% of Corn  71% of Cotton  75% of soybeans

Silk is Soy

Why 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’s  Or Pepsi  Or Coke  Or CapN Crunch???  Or Pepsi  Or Coke  Or CapN Crunch???

How to Manipulate DNA in the Lab

Examples of Transformation  Natural Systems  Bacteria  Viruses  Natural Systems  Bacteria  Viruses

Bacterium bacterial chromosome plasmid

Transformation with DNA fragment bacterial chromosome DNA fragments

Virus enters host cell. 2 virus viral DNA host cell host cell DNA “hybrid virus” viral proteins viral DNA Virus attaches to susceptible host cell. 1 Virus releases its DNA into host cell; some viral DNA (red) may be incorporated into the host cell’s DNA (blue). 3 Viral genes encode synthesis Of viral proteins and viral gene Replication. Some host cell DNA May attach to replicated viral DNA (red/blue). 4 New viruses assemble; host cell DNA is carried by “hybrid viruses.” 5 Host cell bursts, releasing newly assembled viruses. when “hybrid viruses” infect a second cell, they may transfer genes from the first cell to the second cell. 6

GE 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 Based on the Central Dogma and the fact that in virtually organisms the CODE is perfectly conserved almost

Amplifying 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 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 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 polymerases assemble new DNA strands Fig. 16-6, p. 256 Stepped Art

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 Fingerprinting Guilty or Innocent

8 side-by-side (tandem) repeats of 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 power supply DNA bands (not yet visible) �� wells pipetter nylon paper DNA samples are pipetted into wells (shallow slots) in the gel. Electrical current is sent through the gel (negative at end with wells, positive at opposite end). Electrical current moves DNA segments through the gel. Smaller pieces of DNA move farther toward the positive electrode. Gel is placed on special nylon paper. Electrical current drives DNA out of gel onto nylon. ��

gel power supply �� wells pipetter DNA samples are pipetted into wells (shallow slots) in the gel. Electrical current is sent through the gel (negative at end with wells, positive at opposite end). ��

DNA bands (not yet visible) �� Electrical current moves DNA segments through the gel. Smaller pieces of DNA move farther toward the positive electrode. ��

gel �� nylon paper Gel is placed on special nylon paper. Electrical current drives DNA 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 from 13 different people Number of repeats

STR name Penta D CSF D16 D7 D13 D5 Number of repeats

D16: an STR on chromosome 16 DNA samples from 13 different people Number of repeats

Genetic Engineering Transformation 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

Process 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 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.

Discussion  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 judge  With your pocket book  Or your advocacy  With your pocket book  Or your advocacy

Ethical 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?