BIOL 1210 Lecture Notes - Lecture 13: Enzyme Inhibitor, Herbivore, Chief Operating Officer
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View/perform/read ALL THREE of the following prior to answeringthe questions.
http://highered.mcgraw-hill.com/olcweb/cgi/pluginpop.cgi?it=swf::535::535::/sites/dl/free/0072437316/120078/micro10.swf::Stepsin Cloning a Gene (Links to an external site.)
http://www.discoverbiotech.com/wiki/-/wiki/Main/Applications ofCloning (Links to an external site.)
http://www.wiley.com/college/boyer/0470003790/animations/cloning/cloning.htm(Links to an external site.)
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From the list below, which of the following is the most logicalsequence of steps for splicing foreign DNA into a plasmid andinserting the plasmid into a bacterium?
I. Transformbacteria with recombinant DNA molecule
II. Cutthe plasmid DNA using restriction enzymes
III. Extractplasmid DNA from bacterial cells
IV. Hydrogen-bondthe plasmid DNA to nonplasmid DNA fragments
V. Useligase to seal plasmid DNA to nonplasmid DNA
| IV, V, I, II, III |
| III, II, IV, V, I |
| III, IV, V, I, II |
| II, III, V, IV, I |
| I, II, IV, III, V |
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Plasmids (or vectors) are important in biotechnology becausethey are
| a vehicle for the insertion of recombinant DNA intobacteria. |
| surfaces for respiratory processes in bacteria. |
| recognition sites on recombinant DNA strands. |
| surfaces for protein synthesis in eukaryotic recombinants. |
| proviruses incorporated into the host DNA |
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Plasmids are put into bacterial cells by
| restriction enzymes |
| DNA ligase |
| binding of cohesive sticky ends |
| transformation |
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Restriction enzymes usually
| cut donor DNA evenly so smooth edges result |
| cut donor DNA but do not affect plasmids |
| make staggered cuts at specific sequences in DNA in both donorDNA and plasmid |
| are used in ligating plasmids into bacterial host cells |
| more than one of the above |
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After combining DNA fragments in a cloning experiment, ___ isused to covalently join the DNA segments.
| Restriction enzyme |
| DNA Ligase |
| Reverse transcriptase |
| DNA polymerase |
| Helicase |
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It is theoretically possible for a gene from any organism tofunction in any other organism. Why is this possible?
| All organisms have ribosomes. |
| All organisms have the same genetic code. |
| All organisms are made up of cells. |
| All organisms have similar nuclei. |
| All organisms have transfer RNA. |
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Assume that you are trying to insert a gene into a plasmid andsomeone gives you a DNA sample cut with restriction enzyme X. Thegene you wish to insert from the given sample has sites on bothends for cutting by restriction enzyme Y. You have a plasmid with asingle site for Y, but not for X. Your strategy should be to
| cut the plasmid with restriction enzyme X and insert thefragments cut with Y into the plasmid. |
| cut the plasmid with enzyme X and then insert the gene into theplasmid. |
| cut the DNA again with restriction enzyme Y and insert thesefragments into the plasmid cut with the same enzyme. |
| cut the plasmid twice with restriction enzyme Y and ligate thetwo fragments onto the ends of the human DNA fragments cut withrestriction enzyme X. |
| insert the fragments cut with X directly into the plasmidwithout cutting the plasmid. |
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Which of the following is/are false in regard to expressionplasmids (also called expression vectors)?
| They are used to make proteins using a cloned gene. |
| They contain a promotor. |
| They are the first plasmid type used to clone a gene. |
| They contain a terminator. |
| More than one of the above is false. |
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What is NOT a potential problem(s) associated with usingbacteria containing a cloned eukaryotic gene (e.g. a human gene) toproduce a functional protein?
| If the eukaryotic gene contains introns the bacteria will notremove them and the resulting amino acid sequence will be differentthat that made by a eukaryote. |
| The bacteria may not fold the protein correctly. |
| The bacteria may degrade the protein. |
| All of the above are potential problems. |
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Cloning allows for production of proteins in much larger amountsthan occurs in the cells from which the gene is isolated.
| True |
| False |
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Question 111 pts
Gene cloning is used to do all of the following except
| Make insulin |
| Making genetically identical animals (e.g. Dolly thesheep) |
| Make vaccines |
| Perform Gene Therapy |
| Making genetically engineered plants |
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In your
QUESTION 10
If you can drink milk as an adult, it means that you have inherited a mutation in the promoter of your lactase gene (the gene that encodes the enzyme you need to break down lactose). Predict the effect of this mutation:
| The mutation changes the number of domains in the enzyme, which makes it work more efficiently | ||
| The mutation changes the amino acid sequence of the lactase protein | ||
| The mutation increases the number of copies of the lactase gene that will be found in your genome | ||
| The mutation changes whether the lactase sequence is found in an intron or exon | ||
| The mutation affects the expression of the lactase gene |
1.2 points
QUESTION 11
A competitive inhibitor is decreasing the activity of an enzyme. Predict the effect of adding more substrate to the reaction.
| The substrate will increase the reaction rate by binding to the allosteric site | ||
| The substrate will increase the reaction rate by competing with the inhibitor for the active site | ||
| The reaction rate will not change unless the inhibitor can be removed | ||
| The enzyme adjusts its shape so that the substrate, but NOT the competitive inhibitor, can bind | ||
| The substrate will bind to the competitive inhibitor and block its ability to bind to the enzyme |
1.2 points
QUESTION 12
What determines where in the genome a transcription regulator will bind?
| Transcription regulators bind to the 5' UTR region of a gene | ||
| Regulators bind via complementary base-pairing to certain DNA molecules | ||
| Covalent bonds form between the transcription regulator and the atoms of the DNA backbone | ||
| Every eukaryotic gene has a different transcription regulator that will bind to the 5' end of the gene | ||
| Transcription regulators bind to specific DNA sequences via multiple weak non-covalent interactions |
1.2 points
QUESTION 13
What is the basic premise of cell theory?
| DNA -> RNA -> protein | ||
| All cells arise from pre-existing cells | ||
| DNA provides the complete instructions to create a cell | ||
| The identity of a cell is determined through gene expression patterns | ||
| All cells contain the same four basic macromolecules |
1.2 points
QUESTION 14
What is the benefit of using BOTH the lac activator and the lac repressor to control gene expression?
| Using both an activator and repressor enables cells to more accurately determine the amount of lactose available in the environment | ||
| Enzymes to digest lactose are only made when energy is low and lactose is available | ||
| The activator can override the inhibition of the lac operon by the repressor | ||
| The repressor can control the enhancer, while the activator can control the promoter | ||
| When neither the lac activator or repressor is present, expression of the lac operon is too high |
1.2 points
QUESTION 15
What is the histone code used for?
| Phosphorylation and acetylation of DNA affect its ability to be compacted | ||
| Changes to the sequence of DNA change whether DNA will wrap around histone proteins | ||
| Covalent modifications of histones affect the ability of the transcription initiation complex to form | ||
| Histones provide the codon sequences needed for translation to occur | ||
| The histone code affects which amino acids will get added to a polypeptide |
