Biotechnology  Biotechnology involves human manipulation of the genetic code.  Genetic engineering is the process of manipulating genes for practical purposes.  Recombinant DNA-DNA from two or more organisms.  Today we use genetic engineering to produce the protein hormone insulin.  The human gene for insulin is placed in a bacteria and the bacteria will grow the insulin.

Making Recombinant DNA  Step 1:Cutting DNA- DNA of the gene of interest and the vector is cut by a restrictive enzyme. A restrictive enzyme will cut DNA at a specific point between nucleotides. A restrictive enzyme will cut DNA at a specific point between nucleotides. A vector carries the segment of DNA to a cell. A vector can be a virus, yeast, or a plasmid. A vector carries the segment of DNA to a cell. A vector can be a virus, yeast, or a plasmid. Plasmids are pieces of circular DNA that can replicate independently of the main chromosomes of the bacteria. Plasmids are pieces of circular DNA that can replicate independently of the main chromosomes of the bacteria.

Making Recombinant DNA  Step 2: Making recombinant DNA- The DNA segment that carries the gene of interest is combined with the vector.  The enzyme DNA ligase is added to help the DNA of the gene of interest stick to the vector DNA. The recombinant DNA is taken up by the host (bacteria)

Making Recombinant DNA  Step 3: Cloning- When the host bacteria reproduces by binary fission, the gene of interest in the plasmid will be copied as well.  Step 4: Screening- Cells that have received the gene we want are separated out. The bacteria cells will transcribe and translate the protein coded for by the gene.

Transgenic Organism  Animals that have recombinant human DNA that can produce medically important proteins. Bacteria used to make human insulin Bacteria used to make human insulin Pigs used to make human hemoglobin Pigs used to make human hemoglobin

Polymerase Chain Reaction  PCR is and instrument used to replicate a large amount of DNA from a small sample  Lab technique that recreates the conditions of the S-Phase (DNA Replication) of the cell cycle.  DNA to be copied, Polymerase, free bases (A,T,G,C), and Primers are added to the PCR.

Polymerase Chain Reaction  1. Raise the temperature to break hydrogen bonds of DNA, separating the strands to reveal unpaired bases.  2. Lower the temperature to prevent DNA from rebinding. Primers are added.  3. Raise the temperature subtly to an optimum range for Polymerase to add free nucleotides to their complements.  DNA is doubled every 2 minutes. In a couple of minutes millions of copies can be made.   molecularbiology/pcr.html molecularbiology/pcr.html molecularbiology/pcr.html

DNA Fingerprinting  DNA in the nucleus of a human cell is 6ft. long with 3 billion base pairs.  Human DNA has 30,000 places that code for proteins- genes  Each gene is 10,000 base pairs long  Only 10% of our entire 3 billion base pairs code for proteins  90% of out DNA is Non Coding DNA  All humans have 99% of our genes in common.

DNA Fingerprinting  Differences in human DNA is due to non coding regions between our genes.  Variability is due to repeated base sequences  RFLP-Restriction Fragment Length Polymorphisms  Because of these repeated base sequences we can separate DNA fragments out by size using Gel Electrophoresis

Gel Electrophoresis  A large amount of DNA is replicated by PCR.  DNA Fragments are cut by Restriction Enzyme  DNA strands of varying length are placed in a gel that has been treated with a dye that binds to DNA and glows in UV light.  The gel is then subjected to an electrical current carried by a buffer solution.

Gel Electrophoresis  The DNA strands move to the opposite end of the chamber, but at different speeds  Since DNA is negatively charged it will move to the positive pole and the smallest DNA fragments will move the fastest.  A pattern of bands is formed that can be compared to other bands of interest.  The bands can be stained or observed under UV light to determine similarities between band patterns.

Why?  DNA Fingerprinting DNA left at a scene DNA left at a scene Determining the identity of remains Paternity Tests Determining the identity of remains Paternity Tests  bioac/genetics/Electro.htm bioac/genetics/Electro.htm bioac/genetics/Electro.htm  nisamples/majorsbiology/gelelectrophoresi s.html nisamples/majorsbiology/gelelectrophoresi s.html nisamples/majorsbiology/gelelectrophoresi s.html

Cloning  Creation of an organism that is an exact copy of another (same DNA)  Artificial Twinning  Somatic Cell Nuclear Transfer Somatic Cell- body or non reproductive cell Somatic Cell- body or non reproductive cell Nuclear- nucleus that contains DNA Nuclear- nucleus that contains DNA Transfer- to move Transfer- to move  /whatiscloning/ /whatiscloning/ /whatiscloning/

Artificial Embryo Twinning  Manually recreates what occurs when natural clones (identical twins) come about. Single sperm fertilizes a single egg. Single sperm fertilizes a single egg. During early cell division a split occurs leading toward two separate and genetically identical cell masses. During early cell division a split occurs leading toward two separate and genetically identical cell masses.

Somatic Cell Nuclear Transfer 1. Obtain Somatic Cell 2. Remove nucleus from the Somatic Cell 3. Isolate a egg cell 4. Remove nucleus from egg cell 5. Insert somatic cell nucleus into egg cell 6. Induce egg cells to divide with electrical current 7. Implant embryo into surrogate mother to be carried to term

Cloning  Problems with Cloning Extremely Low Success Rate Extremely Low Success Rate Clones have been shown to have numerous developmental issues as well as poor immune systems Clones have been shown to have numerous developmental issues as well as poor immune systems

Stem Cells  Stem Cells are undifferentiated cells that will differentiate and divide into specific cells  Every cell in the body “stems” from these cells  Throughout development, Stem Cells have differing ability to differentiate into many types of cells

Stem Cells  Totipotent- Stem cells that can become any type of cells. Derived from eight cell embryo. “Embryonic stem cells”  Pluripotent- Stem cells that can become almost any type of cell. Derived from a Blastocyst- hollow mass of about 100 cells.  Multipotent- Stem cells that can differentiate a limited range of cells. Derived at birth, the umbilical cord, and adults. “Adult Stem Cells”

Human Genome Project  A genome is all of the base pairs of all the DNA of an organism.  The human genome is the identification of all the base pairs that make up the DNA of humans  Researchers identified 6 feet of DNA with 3.2 billion base pairs.  Only 1-1.5% of DNA codes for proteins.  Only about 30,000 genes.