Biotechnology and Genetic Engineering Notes

1. Biotechnology and Genetic Engineering Notes Important for last MP test and FINAL!

2. Gene Therapy • Mutant or nonfunctional genes are replaced by functional genes • This procedure is only done with patients that have a serious genetic diseasewhere they would normally not live a good/normal life.

3. Gene Therapy • Steps of Gene Therapy • Obtain the normal gene from a healthy person and clone it. • Insert the good gene into the DNA of a virus • The virus will be used as a vector (vehicle), which will carry the gene into normal cells when it infects those cells • Allow cells of the patient to be mixed with the virus carrying the normal gene. The virus will enter the normal cells. • Completed on cells that can be removed from the body • The virus DNA, along with the human gene, will be inserted someplace into one of the chromosomes within the cells of the patient. • Patients cells will replicate. DNA will now contain the new normal gene. The viral genes have been altered so it cannot cause infection. • Remember the virus is just used as a VEHICLE to move the genes • Genetically engineered cells are then put back into the patient – should be able to produce the desired protein

4. DNA Fingerprinting • Used to compare the DNA from 2 different places • Crime scene (suspect’s DNA) • Paternity • Differences exist in the “Junk DNA” portion • Because if there are mutations in these DNA regions, they tend to be passed on through the generations (they do not affect the proteins produced)

5. DNA Fingerprinting • Take DNA from cells • DNA in the sample is cut into small fragments with restriction enzymes Note: the plasmid used in cloning should have ONE restriction site (same used on human)

6. DNA Fingerprinting • The fragments taken are then subjected to a process gel electrophoresis • The fragments of DNA are loaded into wells cut into an agar gel. The gel contains tiny pores. The gel is placed into a box, immersed in a solution that conducts an electrical current. The box is hooked up to a power supply. The wells are placed on the side of the box that has the NEGATIVE electrode • DNA is negatively charged due to the PO4- group. • The DNA will move towards the positive end. • As the pieces move, they are separated out by size (because of the pores in the gel) • Shorter pieces travel farther because they can move easily. • The larger pieces remain further behind. • Power is turned off after a certain amount of time and the fragments stop moving.

7. Gel Electrophoresis

9. The DNA gel is then stained – visualize a banding pattern. • Banding pattern from 2 individuals will be different due to the different number of times and places that the restriction enzymes cut their DNA. • Each band represents many pieces of DNA that were the same size.

13. If you do not understand why the previous answer was B… read here • Remember SHORTER fragments move farther • LONGER fragments move the least • Section B is the shortest piece, then A, then C is the longest piece of DNA. • So the gel should come out C A  B • With the correct distance between.

16. Cloning • Take a cell from an organism (body cell) to be cloned and remove the nucleus/DNA • Take an EGG cell and remove DNA • Insert DNA from skin cell and put into the egg cell • Electrical shock / environmental stress causes cell to take up DNA and start dividing. • Allow embryo to grow in vitro to certain size • Implant into host mom to complete development.

18. Cloning Uses • Organisms • Organs • Tissues • Genes (Bacteria)

19. Stem Cells • Can develop into many different cell types in the body during early life and growth • Undifferentiated • Embryonic Stem Cells • Somatic/Adult Stem Cells • Induced Pluripotent Stem Cells