Turner College & Career High School  2017 DNA Technology Turner College & Career High School  2017

Manipulating DNA Ever since humans have been domesticating animals and raising crops they have been (unwittingly) manipulating genes. By cross pollination and cross breeding they have tried to introduce the beneficial characteristics of one variety into a different variety of the same species. This has been done for thousands of years without any knowledge of genes or the mechanism of inheritance.

Genetic Engineering Genetic Engineers can alter the DNA code of living organisms. Selective Breeding Recombinant DNA PCR Gel Electrophoresis Transgenic Organisms

Selective Breeding Breed only those plants or animals with desirable traits. People have been using selective breeding for 1000’s of years with farm crops and domesticated animals.

Recombinant DNA The ability to combine the DNA of one organism with the DNA of another organism. Recombinant DNA technology was first used in the 1970’s with bacteria.

Benefits of Recombinant Bacteria Bacteria can make human insulin or human growth hormone. Bacteria can be engineered to “eat” oil spills.

Altering the DNA of Plants & Animals Disease-resistant and insect-resistant crops. 2. Hardier fruit. 3. 70-75% of food in supermarket is genetically modified.

Creating a Genetically Modified Plant Create recombinant bacteria with desired gene. Allow the bacteria to “infect" the plant cells. Desired gene is inserted into plant chromosomes.

Genetically modified organisms are called transgenic organisms. TRANSGENIC ANIMALS Mice – used to study human immune system Chickens – more resistant to infections Cows – increase milk supply and leaner meat 4. Goats, sheep and pigs – produce human proteins in their milk

Transgenic Goat . This goat contains a human gene that codes for a blood clotting agent. The blood clotting agent can be harvested in the goat’s milk. Human DNA in a Goat Cell

How to Create a Transgenic Animal Desired DNA is added to an egg cell.

Polymerase Chain Reaction PCR PCR allows scientists to make many copies of a piece of DNA. Heat the DNA so it “unzips”. 2. Add the complementary nitrogenous bases. 3. Allow DNA to cool so the complementary strands can “zip” together.

Gel Electrophoresis This technology allows scientists to identify someone’s DNA!

Steps Involved in Gel Electrophoresis 1. “Cut” DNA sample with restriction enzymes. 2. Run the DNA fragments through a gel. 3. Bands will form in the gel. 4. Everyone’s DNA bands are unique and can be used to identify a person. 5. DNA bands are like “genetic fingerprints”.

DNA Extraction Chemical treatments cause cells and nuclei to burst. The DNA is inherently sticky, and can be pulled out of the mixture. This is called “spooling” DNA.

Cutting DNA Restriction enzymes cut DNA at specific sequences. Useful to divide DNA into manageable fragments.

Electrophoresis DNA can be separated based on size and charge. The phosphate groups are negatively charged. DNA is placed in a gel and electricity is run through.

Electrophoresis Negative DNA moves toward the positive end. Smaller fragments move farther and faster.

Electrophoresis

Steps in DNA Sequencing Many copies of a single strand of DNA are placed in a test tube DNA polymerase is added A mixture of nucleotides is added some of which have dye molecules attached Each base (A,T,C,G) has a different color dye

Steps in DNA Sequencing By chance, some dyed nucleotides & some regular ones are added Dye molecules are large and stop the chain from growing

DNA Sequencing The result is DNA fragments of multiple sizes with colors that can be identified

DNA Sequencing After the gel separates the resulting fragments by size, we 'read' the sequence from bottom to top.

Copying DNA Polymerase Chain Reaction Also called PCR A method of making many copies of a piece of DNA

Steps in Copying DNA A DNA molecule is placed in a small test tube DNA polymerase that can work at high temps is added

Steps in Copying DNA The DNA is heated to separate the two strands Primers, short pieces of DNA complementary to the ends of the molecule to be copied, are added

Copying DNA The tube is cooled, and DNA polymerase adds new bases to the separated strands

Large amounts of DNA can be made from a small starting sample PCR Large amounts of DNA can be made from a small starting sample

Cloning Clone- a member of a group of genetically identical cells May be produced by asexual reproduction (mitosis)

Cloning organisms A body cell from one organism and an egg cell from another are fused The resulting cell divides like a normal embryo

Cloning “Dolly”

Human Genome Project

Human Genome Project Started in 1990 Research effort to sequence all of our DNA (46 chromosomes) Over 3.3 billion nucleotides Mapping every gene location (loci) Conducted by scientists around the world

HGP Insights Only 2% of human genome codes for proteins (exons) Other 98% (introns) are non-coding Only about 20,000 to 25,000 genes (expected 100,000) Proteome – organism’s complete set of proteins About 8 million single nucleotide polymorphisms (SNP) – places where humans differ by a single nucleotide About ½ of genome comes from transposons (pieces of DNA that move to different locations on chromosomes)

Benefits of Human Genome Project Improvements in medical prevention of disease, gene therapies, diagnosis techniques … Production of useful protein products for use in medicine, agriculture, bioremediation and pharmaceutical industries. Improved bioinformatics – using computers to help in DNA sequencing …

Benefits of Genetic Engineering

Biotechnology - The use of gene science to create new products from plants and animals

Biotechnology Provides: Improved food products Medical advances An enhanced environment

Herbicide Resistant Crops + CP4 EPSPS = Roundup gene Ready Soybeans: Roundup Ready Corn: Roundup Ready, Liberty Link Cotton: BXN, Roundup Ready Canola: Liberty Link, Roundup Ready

Biotechnology Breakthroughs Insulin (1982) First commercial biotech product Reliable, inexpensive source of insulin Rice Enriched with beta-carotene and iron Bananas Containing edible hepatitis vaccine

Biotechnology Breakthroughs Potatoes with higher solid content Garlic that lowers cholesterol Fruits and vegetables that reduce risks of cancer and heart disease

Environmental Benefits Reduced pesticide use Lower energy requirements Cleaner water Less soil erosion