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Replication Protein /Mutations Synthesis DNA Structure DNA Control Mechanisms Biotechnology 10 10 10 10 10 20 20 20 20 20 30 30 30 30 30 40 40 40 40 40 50 50 50 50 50 Question 1 - 10 What kind of mutation has occurred in the following DNA sequences: Original strand: Mutated strand: 5’ ACT TGA CGA 3’ 5’ ACT TGA GGA 3’ Answer 1 – 10 A point mutation (also called a substitution) has occurred on the third set of nucleotides. If we transcribed the DNA to RNA and then used a codon chart, we could determine what kind of point mutation it is: missense, nonsense, or silent. Question 1 - 20 What kind of mutation has occurred in the following DNA sequences: Original strand: 5’ ACT TGA CGA 3’ Mutated strand: 5’ ACT TGA CCG A 3’ Answer 1 – 20 A frame-shift mutation has occurred; specifically, an insertion (a base was added). If a base was instead deleted, it would also be a type of frame-shift mutation. They both drastically change the code following the insertion or deletion. The message goes from making biological sense to being gibberish. Question 1 - 30 There are three kinds of point mutations—name them in the order given. 1. This point mutation causes the codon to code for a different amino acid. 2. This point mutation causes the codon to code for an early stop codon. 3. This mutation causes the no change to the resulting amino acid. Answer 1 – 30 By definition: Number one is a missense point mutation. Number two is a nonsense point mutation. Number three is a silent point mutation. Question 1 - 40 Which strand, A or B, would be the LAGGING strand. A B Answer 1 – 40 Strand B would be the lagging strand. Strand B would start where the replication fork is and build antiparallel to the original strand, from 5’ to 3’. As it does this, the DNA continues to separate and then there is a gap where polymerization has to start again. This creates [Okazaki] fragments instead of one continuous strand. Question 1 - 50 In DNA replication, name the 4 enzymes that construct a new complimentary DNA strand, in the order of their use. Answer 1 – 50 1. DNA Helicase breaks the hydrogen bonds, separating the two strands 2. Primase lays a RNA primer down where replication will begin. 3. DNA Polymerase (III) adds new nucleotides in the 5’ to 3’ direction starting at the primer. (technically DNA Polymerase (I) also replaces the primer with DNA nucleotides as well) 4. DNA ligase binds the lagging strand fragments together to make a continuous DNA molecule. Question 2 - 10 Using your codon chart, determine the amino acids sequence produced by the following DNA sequence: 3’ TAC ACT 5’ Answer 2 – 10 3’ TAC ACT 5’ 5’ AUG UGA 5’ MET-Stop Question 2 - 20 Using your codon chart, determine the amino acids sequence produced by the following DNA sequence: 5’ GCG TAA 3’ Answer 2 – 20 DNA: 5’ GCG TAA 3’ ----mRNA: 3’ CGC AUU 5’ This is the same as: 5’ UUA CGC 3’ which is: Must be read 5’ to 3’ LEU-ARG Question 2 - 30 Which number corresponds to the following ribosomal sites: A site 1 2 3 E site P site Answer 2 – 30 A site = 3, you can see the tRNA bringing in a new amino acid to be added to the chain E site = 1, the tRNA without an amino acid would be the tRNA molecule exiting P site = 2, the polypeptide is being built here, there are two amino acids bound together Question 2 - 40 What would facilitate the folding of a freshly translated protein if it were to stay in the same cell it was created in? What if it was to be exported from the cell? Answer in order. Answer 2 – 40 Chaperonin facilitates the folding of proteins that stay within the creator cell. The Rough ER facilitates the folding of proteins that are leaving the cell. Question 2 - 50 List the three modifications that must be made to the primary mRNA transcript before it can leave the nucleus. Answer 2 – 50 1. Introns must be removed from the transcript 2. Addition of a 5’ cap 3. Addition of a poly-A tail containing 50-250 adenine based nucleotides Question 3 - 10 If a DNA sample consists of 12 percent Adenine, what percentages would the remaining bases make up? Answer 3 – 10 A= 12%, T= 12%, C= 38%, G = 38% For you mathy folks: A= 12. A=T, therefore T = 12 A + T + C + G = 100. If A + T = 24, then C + G = 76 If C=G, then C= one half of 76. C = 38, G = 38. Question 3 - 20 C B A Identify parts A, B, and C, and state what these three things are called together. Answer 3 – 20 A- Phosphate group B- 5 Carbon Sugar (Pentose Sugar, specifically deoxyribose) C- Nitrogenous Base (this one happens to be adenine) Together they are known as a NUCLEOTIDE. Question 3 - 30 Organize the following items in order from GREATEST to LEAST (use the numbers). 1. Atom 2. Chromosome 3. DNA 4. Gene 5. Genome 6. Nucleotide Answer 3 – 30 The correct order is: 5 , 2, 3, 4, 6, 1 Your genome is all of your genetic material, it is usually made of 46 chromosomes, if you are human. These chromosomes are long strands of DNA plus some other molecules that help it wind. Each DNA molecule is made of many genes. Each gene is made of many nucleotides, and each nucleotide is made of many atoms. Question 3 - 40 What kind of sugar is used in this nucleotide? C 5’ 4’ 1’ 3’ A 2’ B Answer 3 – 40 Deoxyribose There is no oxygen on the 2’ carbon. This is why we call it DEoxy. Question 3 - 50 What did Hershey and Chase use Sulfur-35 and Phosphorus-32 to do in their experiment? What was their conclusion? Answer 3 – 50 Sulfur-35 was used to label the protein coat of the virus. Phosphorus-32 was used to label the phosphate backbone of DNA. When future generations were checked, in the sulfur experiment there was no sign of the radioactive isotope. In the phosphorus experiment, the new generations were still radioactive. This concluded that DNA, not protein, is the molecule of heredity. Question 4 - 10 Which part of the operon, when acted upon by another molecule, turns a gene “on” or “off?” Answer 4 – 10 The operator is the area that can turns a gene on or off by induction or repression. Question 4 - 20 What is a molecule that causes a repressor to unbind with the operator called? Answer 4 – 20 An inducer. Question 4 - 30 What is a molecule that binds with a repressor in order to turn a gene “off” called? Answer 4 – 30 A co-repressor. Question 4 - 40 What is a moveable segment of DNA that is capable of blocking transcription of a gene called? (also known as a “jumping” gene) Answer 4 – 40 A transposon. Question 4 - 50 Can transcription occur given this operon’s current state? Why or why not? Answer 4 – 50 No, because the operator is being inhibited by the repressor molecule. RNA polymerase cannot read the gene and create a mRNA transcript. Question 5 - 10 What is the name of the effort that set out to map the entire human genome called? Answer 5 – 10 The Human Genome Project. Question 5 - 20 What is the name of the proteins that cut DNA into fragments at very specific base sequences? Answer 5 – 20 Restriction enzymes. Question 5 - 30 Which base pair fragment would travel farther, given the same amount of time, in a gel electrophoresis sample: 2000 base-pair fragment 10000 base-pair fragment Answer 5 – 30 The 2000 base pair fragment. It has less mass, and less length, making it easier to navigate through the porous gel. Question 5 - 40 Why do DNA fragments move towards the positive end of a gel electrophoresis chamber when an elecrtical current is sent through the chamber? Answer 5 – 40 Because DNA is a negatively charged molecule. It is repelled by the negative side and attracted to the positive side of the chamber. Question 5 - 50 How are human genes inserted into plasmids to produce a desired protein product? Answer 5 – 50 1. Restriction enzymes are used to produce “sticky ends” on the desired human gene AND the plasmid 2. The sticky ends allow the human gene to be connected to the plasmid and then is introduced to a bacteria sample 3. The bacteria are grown to create the desired protein product (Antibiotic resistance gene is also added to the plasmid to identify which bacterial cells successfully took up the plasmid. Only the ones that take up this plasmid will survive when exposed to antibiotics) Final Jeopardy • Make your wager! • Calculate the rate of the following graph!