Term
| What is genetic engineering? |
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Definition
| The induced mutation of genes to produce a product that can be harvested |
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Term
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Definition
| Creating a genetically identical copy of a organism |
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Term
| What is recombinant DNA technology? |
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Definition
| Creating, cloning and utilising recombinant DNA |
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Term
| What is needed to do recombinant DNA technology? |
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Definition
| DNA/RNA, enzymes, vectors and cells |
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Term
| What are the four types of cell used in recombinant DNA technologies? |
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Definition
| Bacteria, yeast, insect and mammalian |
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Term
| What are the four enzymes used recombinant DNA technologies? |
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Definition
| Restriction enzymes, DNA ligase, Taq polymerase ad Reverse Transcriptase |
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Term
| What do restriction enzymes do? |
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Definition
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Term
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Definition
| Stick fragments of DNA together |
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Term
| What does Taq Polymerase do? |
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Definition
| Alows DNA replication to occur at the high temperatures in PCR |
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Term
| What does reverse transcriptase do? |
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Definition
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Term
| How did restriction enzymes come about? |
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Definition
| Defense mechanism against bacteriophages |
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Term
| How do restriction enzymes know which section of DNA to cut? |
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Definition
| They recognise the palindromic sequences |
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Term
| What is a palindromic sequence? |
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Definition
| A sequence that is the exact copy on the template strand |
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Term
| What are the two types of cut a restriction enzyme can do? |
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Definition
| Symmetrical and asymmetrical cleavage |
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Term
| What does symmetrical cleavage produce? |
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Definition
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Term
| What does asymmetrical cleavage produce? |
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Definition
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Term
| What are the two types of sticky end? |
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Definition
| 5' overhang and 3' overhang |
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Term
| What are the four properties of a good vector? |
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Definition
1) Has unique restriction sites for DNA insertion
2)Is competent
3)Has an origin of replication
4)Contains a gene that produces a selection factor |
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Term
| What is the most common vector? |
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Definition
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Term
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Definition
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Term
| What are the two other vectors? |
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Definition
| Bacteriophages and cosmids |
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Term
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Definition
| A plasmid that can be inserted into bacteriophages |
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Term
| How do we create recombinant DNA? |
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Definition
| Obtain the DNA of interest and insert it into a vector |
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Term
| Where does the DNA in recombinant DNA technologies come from? |
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Definition
| Random fragments of DNA cut up to form a genomic library or a cDNA library |
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Term
| What is a genomic library? |
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Definition
| Lots of fragments of one DNA cut up |
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Term
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Definition
| The copy of DNA using reverse transcriptase |
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Term
| How is the DNA amplified? |
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Definition
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Term
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Definition
| Polymerase Chain reaction |
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Term
| How is the DNA inserted into a vector? |
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Definition
1)Restriction enzyme cuts ope plasmid
2)Gene is then inserted and DNA ligase then seals up the gap |
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Term
| How do we clone recombinant DNA? |
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Definition
| Vector is inserted into a host, host then divides many times, the many vectors are then purified |
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Term
| What are the two ways that a vector can be inserted into a bacteria? |
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Definition
| Heat shock or electroporationj? |
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Term
| How does heat shock work? |
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Definition
| CaCl2 is added over ice, then they are heated to 42 degrees |
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Term
| How does electroporation? |
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Definition
| A series of electric shocks are administered over the membrane to disrupt it |
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Term
| After the vector is inserted what are the three possible types of cell? |
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Definition
->Taken up plasmid
->Taken up recombinant plasmid
->Not taken up any plasmid |
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Term
| How are bacteria selected for recombinant DNA? |
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Definition
| Using ampicillin resistence |
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Term
| What type of plasmid is usually used for vectors? |
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Definition
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Term
| Outline the mechanism of identifying recombinant DNA bacteria with Ampicillin and Tetracyline? |
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Definition
1)Grow on ampicillin kills off those without plasmid
2)Grow on tetracycline kills off recombinant DNA bacteria
3)Then go back to work out which were killed off on original plate |
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Term
| Outline mechanism of identifying recombinant DNA with the LacZ gene? |
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Definition
1)Grow on plate containing X-gal
2)Those with LacZ gene (No recombinant DNA) will glow blue as they will break down X-gal
3)Those without LacZ gene (recombinant DNA present) will not glow as they cant break down X-gal |
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Term
| What are the three ways we can identify a colony containing the cloned DNA? |
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Definition
| Complimentary DNA probes, PCR of primers and screening for the expression of a particular protein |
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Term
| How can we use recombinant DNA? |
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Definition
| To express proteins, manufacture the plasmid or transfer the plasmids |
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Term
| What is an expression vector? |
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Definition
| A plasmid specifically made for expressing proteins |
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Term
| What must an expression vector contain as well as the gene? |
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Definition
| A promoter sequence before the gene |
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Term
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Definition
| Junk DNA between the exons |
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Term
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Definition
| The parts of DNA that make up mRNA |
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Term
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Definition
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Term
| What does pre-mRNA contain? |
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Definition
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Term
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Definition
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Term
| What are the four advantages of using bacterial cells? |
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Definition
1)Simple
2)Fast replication
3)Large yield
4)Low costs |
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Term
| What are the three disadvantages of using bacterial cells? |
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Definition
1)Eukaryotic proteins can't be folded correctly
2)Proteins can be toxic to bacteria
3)No post-translational modification |
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Term
| What are the four advantages of using yeast cells? |
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Definition
1)Unicellular eukaryote
2)Similar to mammals
3)Grows quickly and cheaply
4)Perform post-translational modifications |
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Term
| What are the two disadvantages of using yeast cells? |
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Definition
1)Proteases degrade some proteins
2)What are the four advantages of using bacterial cells? |
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Term
| What are the four advantages of using insect cells? |
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Definition
1)High level protein expression
2)Correct folding of mammalian proteins
3)Cheaper than mammalian cells
4)Post-translational modifications |
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Term
| What is the disadvantage of using insect cells? |
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Definition
| 1)What are the four advantages of using bacterial cells? |
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Term
| What are the three advantages of using mammalian cells? |
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Definition
1)Produce mammalian proteins
2)Fold proteins correctly
3)Post-translational modifications are correct |
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Term
| What are the three disadvantages of using mammalian cells? |
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Definition
1)Complexity
2)Lower cell counts
3)Expensive |
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Term
| Name one product or recombinant DNA technologies used commercially? |
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Definition
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Term
| What are the three problems with using animal insulin? |
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Definition
1)Side effects of it being different
2)Difficult to purify
3) Possible contamination |
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Term
| How has recombinant DNA technology been used in HIV research? |
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Definition
| CD4 antibody blocks CD4 receptor and HIV cannot infect. Adding the recombinant DNA that blocked the production of the CD4 receptor prevented HIV infection |
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