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• What are mutations? • What are Genetic disorders? • Trivia tidbit: A tobacco plant has the ability to glow! How so? • Can humans glow? • What are transgenic organisms? Human Genetics-biotechnology Vocabulary • Recombinant DNA DNA that has been genetically modified by connecting DNA fragments from multiple sources • Host organism you are obtaining the gene from • Vector organism such as a bacteria, you are going to use to put the recombinant DNA into the organism you are trying to change • Plasmid DNA or “chromosome” of a bacteria • Restriction Enzyme bacterial proteins that have the ability to cut both strands of DNA at specific points called restriction sites Biotechnology • Biotechnology the use of technology to manipulate or change life in some manner – The only way to do this is through changing genes somehow • Can occur in many different ways. – – – – – Gene manipulation and DNA recombination Gene Therapy Cloning Stem Cells Selective breeding and hybridization Process of manipulating genes 1. Choose an organism to change, and one to obtain the gene from (host) 2. Choose a bacterial vector that will transform the gene incorporate gene into its own DNA 3. Gene Splicing Cleave (cut) the pieces of DNA of interest from the host organism restriction enzyme“cleaves” 4. Isolate the gene located in the pieces of DNA in the host 5. Insert the gene into bacterial vector DNA (recombinant DNA) 6. Expose the vector to the organism you are trying to change 7. Organism must “take up” recombinant DNA Crystal Jellyfish Genetic Engineering • Scientists alter genes of an organism – Usually done by Inserting DNA from one organism into another – DNA that has been genetically modified is called recombinant DNA – Organisms with recombinant DNA are called genetically modified organisms (GMO) or transgenic organisms Using Restriction enzymes to make recombinant DNA • Restriction enzymes are chemical catalysts that “cut” DNA molecules at specific locations in order to obtain certain genes – Can be used for genetic manipulation or DNA cloning • Each Restriction enzyme is specific to a specific restriction site • They are cut into a specific pattern allowing there to be sticky ends – These sticky ends can form hydrogen bonds with other sticky ends creating a whole piece of DNA with the potenetial for them being from different sources Cloning a Eukaryotic gene in a bacterial plasmid • Works the same was as making recombinant DNA with one exception: • You obtain a piece of DNA from a bacteria, this is called the cloning vector – Use a restriction enzyme to produce sticky ends • You obtain genes from a human cell • You then expose the human pieces of DNA into the bacterial cell • Use the enzyme DNA ligase to “glue” the pieces back together • Then you place the bacterial DNA back into the bacteria Transgenic organisms • http://www.youtube.com/watch?v=ujZHrR1m ro8&feature=player_detailpage Applications for gene manipulation • Transgenic bacteria in agriculture • Transgenic bacteria in Industry • Transgenic bacteria in Medicine • Genetically engineered food • Transgenic plants and animals Transgenic bacteria in agriculture • Once species of bacteria have been used successfully on agricultural crops already • Specific bacteria naturally occurs on strawberry plants that promotes frost damage in strawberries because bacteria has ice crystals that form around specific protein – Scientists have been able to isolate the gene for this protein, and prevent frost damage – This saves farmers money, provides jobs to scientists who research and develop this type of transgenic bacteria, and provides higher quality and quantity of strawberries Papaya engineering • Papayas in Hawaii were threatened by papaya ringspot virus. • Scientists in Hawaii isolated the gene in the virus that makes it immune to itself, then implanted that gene into new papaya seeds, and allowed the seeds to reproduce creating new papaya trees that are resistant to the papaya ringspot virus • 1999, the first successful papaya tree produced, now, ¾ of all Hawaiian papayas are genetically modified • Cool fruits: – pluot – Lemato – Tangelo – Diabetes fighting lettuce – Colorful carrots • We have the technology to engineer almost any food Transgenic bacteria in Industry • Scientists have been able to engineer bacteria to be able to break down harmful pollutants into harmless chemicals – Helps environment and provides jobs • Ex: There is a specific bacteria that naturally breaks down oil based compounds. – Scientists genetically engineered bacteria to break down oil at a much faster pace, and with better results Transgenic bacteria in medicine (“Pharming”) • Pharmaceutical companies have developed the widespread use of specific molecules-made by transgenic bacteria that help to treat human diseases • Ex: Human Growth Hormone-given to individuals suffering from dwarfism – Chemotherapy drug Interferon to treat cancer – Insulin to treat diabetes – Artificial sweeteners are usually made from transgenic bacteria that produce phenylalanine – Many treatments for genetic diseases re now being produced by genetic engineering Genetically modified foods • http://www.youtube.com/watch?feature=play er_detailpage&v=FTfuAbzAeB8 Transgenic plants • Glowing tobacco • Plants and animals are much harder to engineer, because they do not contain plasmids What are they again? • Plants – Difficult to engineer, due to thick cell wall – Must use artificial vectors such as a gene gun or a micropipette in other words, they must be done in a lab and not in nature – Main purpose to be able to resist herbicides, produce their own pesticides, or to increase their crop production Blue roses Transgenic animals • Animals usually will not accept a bacterial plasmid as a vector, so manual engineering with a gene gun or a pipette are used when possible • Manipulated genes are inserted into unfertilized eggs • Egg gets fertilized and transgenic traits or properties are expressed • Uses with livestock-placing human proteins into livestock so they produce milk with these proteins already in them – Ex: There is a protein that dissolves blood clots transgenic goats have the protein in their milk, so if person drinks milk, they can dissolve blood clots • Not very far in development – Still a very young field – Ethical issues Green Florescence protein Human Genome Project • In 2000 scientists from 6 countries all worked together and figured out the “book of life” • Wanted to “map out” all nucleotides or bases that give the template for a human • Achieved a blueprint of the whole sequence of bases for humans • Genomics study of entire genomes – Study of all bases in an entire organism Surprise findings • Humans have few genes – Scientists thought 120,000 genes – Reality25,000 genes • Most DNA is non-coding less than 2% actually codes for traits • We share majority of our DNA with other species – 50% same as a fly, 75% same as dog, 30% same as banana, and 98% same as chimp • We all share about 99.9% of our DNA with each other Manipulating bodies and development • In lab=Cloning process used to create identical DNA fragments, cells, or whole organisms • In nature=asexual reproduction Cloning • When you clone an old organism, the newly formed organism’s DNA will have the same age as the organism. – New organism has all the mutations that have occurred in old organism – Ex: Baby sheep would have old DNA but still be young and a baby – Effects: • • • • • • Increased aging rate Increased mutations Shorter life span More genetic issues Abnormal development Ethical issues Four steps to cloning 1. fragmentation - breaking apart a strand of DNA 2. ligation - gluing together pieces of DNA in a desired sequence 3. transformation - inserting the newly formed pieces of DNA into cells 4. screening/selection - selecting out the cells that were successfully transferred with the new DNA Stem cells • 3 types: – Totipotent-can become any type of cell – Pluripotent-can be anything but a sex cell – Multipotent-can be just a few other types • Ex: bone marrow cells • Adult stem cells cells are removed, and used to grow more cells of specific tissue types – Used to replace damaged tissue • Embryonic stem cells-cells from embryos have potential to become all different types of cells Uses for stem cells • To help aid in some cancers such as ones that are tumor related • Transplants • Genetic engineering • Organ failure • Embryonic almost anything Steps for creating stem cells Selective breeding • The method of artificially seelecting and breeding only organisms with a desired trait to produce the next generation. • In other words, you select mates based on the traits you want – Ex: crops, domesticated animals • Once the breeder has successfully produced offspring with the desired set of characteristics, inbreeding will occur Hybridization • Another form of selective breeding to choose and breed organisms that show strong expression for two different traits in order to produce the offspring that express both traits. • This often occurs between two different (but similar) species. The offspring are often healthier than the parents • Ex: Mule, killer bees, Zonkey, Zorse, Liger, Jaglion • In nature polar bear-grizzly bear hybrid with DNA confirmation DNA fingerprinting or gene mapping •Method of comparing different strands of DNA with those of other species or with other people •Method used for formal pregnancy test, crime investigations, and other forensics Gel electrophoresis Gene Map