Sample Examination Questions for Exam 2 Material Biology 3300 / Dr. Jerald Hendrix Warning! These questions are posted solely to provide examples of past test questions. There is no guarantee that any of these questions will be on any examination in the future. Students are responsible for all of the material covered in lectures, assigned readings, textbook problems, laboratories, and any other assigned work. Since these samples have been taken from several past exams, some questions may be very similar or identical. On short answer, essay questions, and genetics problems, the point values from previous exams have been included to give an indication of approximately how much “weight” was given to a question in the past; however, there is no guarantee that any particular question, format, or point distribution will be used on any examination. The following information pertains to questions 3 and 4. The two strands of a double-stranded DNA molecule can be separated by heating the strands. The temperature at which the strands separate is called the melting point of the DNA. A DNA molecule with a larger number of hydrogen bonds between the strands will have a higher melting point than a DNA molecule with fewer hydrogen bonds. In questions 3 and 4: A = the number of deoxyadenosine nucleotides in a DNA molecule C = the number of deoxycytidine nucleotides in a DNA molecule G = the number of deoxyguanosine nucleotides in a DNA molecule T = the number of deoxythymidine nucleotides in a DNA molecule 3. The value of (G + C)/(A + T) (a) (b) (c) (d) (e) is equal to one if the DNA is single-stranded. is equal to one is the DNA is double-stranded. is larger in DNA samples having a higher melting point. is lower in DNA samples having a higher melting point. none of the above. GC base pairs have 3 hydrogen bonds; AT base pairs have only 2. Therefore, if a sample of DNA has more GC, then we expect it to have a higher melting point. 4. The value of (A + C)/(G + T) (a) (b) (c) (d) (e) is equal to one if the DNA is single-stranded. is equal to one is the DNA is double-stranded. is larger in DNA samples having a higher melting point. is lower in DNA samples having a higher melting point. none of the above. In double-stranded DNA, the amount of A equals T, and C equals G, so (A+C)/(G+T) equals one. Page 1 6. Which of the following enzymes is used to cleave vector and chromosomal DNA in the formation of recombinant DNA molecules? (a) (b) (c) (d) (e) 7. In a recombinant DNA expression library (a) (b) (c) (d) 10. trisomy monosomy tetrasomy chromosomal deletion chromosomal translocation A bacterial enzyme that recognizes a specific nucleotide sequence on doublestranded DNA, then cleaves the DNA at the recognition site, is called a (a) (b) (c) (d) (e) 16. the cloned DNA is inserted into an RNA vector to facilitate expression. the cloned DNA is inserted into a plasmid vector at a position adjacent to a bacterial promoter region to allow for transcription and subsequent gene expression. the cloned DNA is bound directly to the small ribosome subunit which results in expression in about 85% of the colonies isolated. the cloned DNA is not expressed inside the bacterial cell. A human being with a chromosome number of 47 in each somatic cell has which of the following conditions? (a) (b) (c) (d) (e) 12. RNA polymerase aminoacyl tRNA synthetase Type I restriction endonuclease Type II restriction endonuclease reverse transcriptase Type I restriction endonuclease. Type II restriction endonuclease. modification methylase. DNA polymerase. 5' exonuclease. The map distance calculated in a two-factor test cross is most likely to be an underestimate of the actual distance due to (a) (b) (c) (d) (e) epistasis. nondisjunction. interference. codominance. double crossovers. Page 2 17. Which of the following enzymes can be used to synthesize cDNA from an mRNA template? (a) (b) (c) (d) (e) 23. DNA-directed RNA polymerase DNA-directed DNA polymerase restriction endonuclease DNA ligase reverse transcriptase The oocytes of the frog Xenopus contain lampbrush chromosomes. These are large, visible chromosomes with both condensed regions and with greatly extended regions that resemble the bristles on a lampbrush. If Xenopus oocytes are incubated with radiolabeled uridine for a brief period of time and autoradiographed, only the extended regions are radiolabeled. What does this indicate? (a) (b) (c) (d) (e) DNA is replicated repeatedly without mitosis in the extended regions. Protein synthesis is occurring in the extended regions. Transcription is occurring in the extended regions. Crossing-over is occurring in the extended regions. Assortment is occurring in the extended regions. Uridine is found in RNA, so radiolabeled uridine would serve as a label for transcripition, or RNA synthesis. 25. Which of the following terms best describes a ribose molecule with a cytosine molecule attached to its 1' carbon and an -OH group attached to its 5' carbon? (a) (b) (c) (d) (e) 31. a nitrogenous base a nucleoside a nucleotide a base pair a promoter In a karyotype of a man whose wife has had several miscarriages, it was discovered that he had only 45 chromosomes. However, he displayed no obvious phenotypic defects. Furthermore, one of the chromosomes of pair 15 was abnormally long. Which of the following terms best describes the condition of this man? (a) (b) (c) (d) (e) monosomy trisomy tetrasomy translocation carrier cri-du-chat Page 3 32. Which of the following terms best describes the procedure in which DNA fragments are separated on an agarose gel, transferred onto a nylon membrane, and screened with a labeled hybridization probe? (a) (b) (c) (d) (e) 1. dot blotting dash blotting Southern blotting Northern blotting Western blotting In the tomato plant, round fruit shape (R) is dominant over long shape (r). Smooth fruit skin (P) is dominant over peachy skin (p). The genes for fruit shape and fruit texture are linked on the same chromosome. A heterozygous round, heterozygous smooth plant was crossed with a long, peachy plant. The results are given in the table below. Smooth round Smooth long Peachy round Peachy long 39 463 451 47 (a) In the heterozygous parent plant, was the linkage between the genes cis or trans? (2 pt) trans (b) Calculate the map distance between the genes. (2 pt) 8.6 map units (c) Is the distance calculated in (b) more likely to be an overestimate or an underestimate of the true distance between the genes? Briefly explain your answer. (4 pt) An underestimate, due to the possibility of double crossovers 2. If the value of interference in a three factor testcross is negative, what does this signify with respect to the process of recombination between the chromosomes in the homologous pair? (4 pt) A negative interference value indicates that the first crossover has somehow increased the probability of a second crossover occurring. 5. How can chromosomal nondisjunction during meiosis result in an offspring with trisomy or monosomy? (4 pt) Nondisjunction is the failure of homologous chromosomes to separate during meiosis I, or sister chromatids to separate during meiosis II. The result is an abnormal gamete that either contains two copies of a chromosome instead of one, or doesn’t contain a copy at all. If an abnormal gamete containing two copies of a chromosome is fertilized with a normal gamete containing one copy, the result is a zygote with a trisomy for that chromsome. If an abnormal gamete missing a chromosome is fertilized with a normal gamete, the result is a monosomy. Page 4 6. Briefly distinguish between trisomy Down’s syndrome and translocation Down’s syndrome. (4 pt) In trisomy Downs syndrome, the individual has three copies of chromosome 21, for a total of 47 chromosomes. In translocation Downs syndrome, the individual has a total of 46 chromosomes, with two normal 21 chromosomes, one normal 14, and an abnormal 14/21. These individuals would be both show the phenotypes for Downs syndrome: retardation, heart problems, facial effects, etc. A translocation Downs carrier shows no symptoms of Downs syndrome, but often will have children with the syndrome. The translocation carrier has 45 chromosomes, with one normal 21, one normal 14, and one abnormal 14/21. 7. What are polytene chromosomes? How are they formed in the cells of Drosophila? What feature of polytene chromosomes suggests that the chromatin of the interphase nucleus is folded in an ordered fashion? (6 pt) Not covered. Page 5 3. In maize, the gene for colored kernels (C) is dominant over its recessive allele for colorless kernels (c). The gene for full kernels (Sh) is dominant over its allele for shrunken kernels (sh). The gene for nonwaxy endosperm (Wx) is dominant over its allele for waxy endosperm (wx). These three genes are located on the same chromosome. A plant heterozygous for all three genes was crossed with a plant that was homozygous recessive for all three genes. The progeny are given in the table below. colored colored colored colored colorless colorless colorless colorless shrunken shrunken full full shrunken shrunken full full waxy nonwaxy waxy nonwaxy waxy nonwaxy waxy nonwaxy 305 112 74 22 18 66 128 275 (a) Determine the sequence of these three genes. (2 pt) Sh – C – Wx (b) State the type of linkage (cis or trans) that existed between each pair of genes in the heterozygous parent. (3 pt) Sh – C: trans; C – Wx: trans; Sh – Wx: cis (c) Calculate the map distance between each pair of genes. (4 pt) Wx – C: 28 map units Sh – C: 18 map units (d) Calculate the coefficient of coincidence and the interference of the cross. (4 pt) Coefficient of coincidence = 0.794 Interference = 0.206 10. A geneticist is studying the structure of two different viruses, A and B, each of which contains DNA. The nucleotide base composition of each virus is given in the table below. Base Adenine Guanine Cytosine Thymine Amount of the base in Virus A Virus B 10000 7000 15000 18000 15000 10000 10000 15000 Based on this data, what differences do you predict will exist between the structures of these two viruses? (4 pt) Virus A most likely has double-stranded DNA, but virus B has single-stranded DNA. Page 6 11. Give two reasons why type II restriction endonucleases are useful in preparing recombinant DNA molecules. In your answer, you should contrast type II restriction endonucleases with type I restriction endonucleases. (4 pt) 1. Type II restriction endonucleases recognize specific nucleotide sequences and cut the DNA at the recognition site, making reproducible fragments. This is different from type I enzymes, because type I enzymes cut DNA at a random distance from teh recognition site and make random fragments, so they are not useful for recombinant DNA technology. 2. Many Type II restriction endonucleases have palindromic recognition sequences and make staggered cuts, so they make short single-stranded complementary ends (“sticky ends”). This means that if DNA from two different sources is cut with the same Type II enzyme, the ends can be spliced together (the DNA is sealed with another enzyme called DNA ligase), making recombinant DNA. 1. Give the correct name for each of the following structures. (4 pt each) O H HO NH 2 H H3C O N C H N O HO O H N O C H P O N OH (a) OH H deoxythymidine (b) O N (c) N N H guanine OH H deoxycytidine monophosphate H N H HO C H O NH 2 OH (d) ribose Page 7 OH OH O O NH2 O N H HO C H O N HO O P H N H O C H O N O OH (e) OH H deoxycytidine 2. Briefly describe the B-helix structure of double-stranded DNA. Include its length in Angstroms (Å) per turn, the number of bases per turn, its grooves, its basepairing, and the orientation of the strands with respect to each other. (10 pt) (f) OH OH uridine monophosphate The sugar-phosphate backbones of the strands are wound around the axis of the helix, forming two grooves that spiral around the axis: a large groove, or major groove, and a smaller groove, or minor groove. The bases are stacked in the center of the helix, where they pair according to the base pairing rules: A to T, with two hydrogen bonds, and C to G with three hydrogen bonds. The helix is 20 Å in diameter, 10 bases per turn, 34 Å per turn, 3.4 Å per base. The strands are antiparallel, meaning that one strand is in the 5’ → 3’ orientation, and the other strand is 3’ → 5’. 3. Briefly explain the two major lines of evidence used by Watson and Crick to deduce the structure of the B-DNA helix. (6 pt) X-ray diffraction crystallography of DNA fibers showed that DNA was in the form of a helix, and from the diffraction pattern Watson and Crick were able to calclulate the dimensions of the helix. Chargaff’s data on base composition of DNA showed that the A:T and C:G molar ratios were 1:1, suggesting that the DNA was a double helix with A-T and C-G base pairs. 5. What is bacterial competence? (2 pt) Competence is the ability of bacterial cells to take up isolated DNA molecules by transformation. Some bacteria are naturally competent for transformation, but other bacteria, such as E. coli, must have competence induced by treatment with calcium chloride (CaCl2) Page 8 12. You wish to study the gene in humans for the protein actin. You know that this protein is made in cultured human fibroblast cells, but it is not the only protein made in the cells. You also know the amino acid sequence of human actin. At the time you are doing your experiment, no actin genes from other species have been isolated. You have available to you a human recombinant DNA library, cultured fibroblasts, purified antibodies specific for the protein actin, and all the supplies and equipment needed to perform the techniques that we discussed in class. The recombinant library is not an expression library. Outline two different experimental approaches to isolate the actin gene from the recombinant library. (14 pt) Since the library is not an expression library, you’ll need a probe to be able to screen the colonies in your library for the actin gene. Where can you obtain your probe? There are two ways we could do this: 1. Isolate the mRNA for actin from cultured human fibroblast cells, and use the mRNA to make a cDNA probe using reverse transcriptase. 2. Create a synthetic DNA probe by deducing possible sequences of the actin gene using the known amino acid sequence of the protein actin and the genetic code table. You can’t use a gene from an existing species since, according to the problem, non exist at the time you are doing your work. Also, you cannot screen the colonies in the library for expression since it is not an expression library (i.e., the fragments are not cloned in an expression vector). Page 9 13. Sickle-cell anemia is caused by a mutation in the human β-globin gene. The three possible genotypes are homozygous for normal β -globin, heterozygous carrier (having both the normal and sickle-cell genes), and homozygous for sickle cell anemia. Recombinant DNA technology has been used as the basis for prenatal diagnosis of sickle cell anemia. In a very high percentage of the cases observed, the normal human β-globin gene is carried on a 7600 bp human DNA fragment from a HpaI digest, while the sickle-cell gene is carried on a 13000 bp HpaI fragment. HpaI is a type II restriction endonuclease. You have available to you: • A radiolabeled sample of recombinant DNA consisting of a bacterial plasmid vector (4000 bp) carrying the 7600 bp HpaI fragment from the normal human genome. • A sample of chromosomal DNA from each member of a couple thought to be carriers of the sickle-cell trait and expecting their first child. • A sample of the chromosomal DNA obtained from the fetal cells in the amniotic fluid from the uterus of the pregnant woman. • All of the supplies and equipment necessary to perform the techniques that we discussed in class. How can you tell if the parents are heterozygous carriers of the sickle-cell trait, and if the fetus is normal, a heterozygous carrier, or homozygous for sickle-cell anemia? (12 pt) A helpful hint: the genes for normal and sickle-cell β-globin share sufficient sequence homology to hybridize with each other. For each parent’s DNA and the fetal DNA, cut the DNA with HpaI and separate the fragments by agarose gel electrophoresis. Using the Southern blotting technique, transfer the fragments to a nylon (or nitrocellulose) membrane, and probe the membrane with the radiolabeled 7600 HpaI fragment. Expected results: Homozygous normal β-globin: Homozygous sickle cell anemia: Heterozygous sickle cell carrier: 11. A single band at 7600 bp A single band at 13000 bp Two bands, one at 7600 and one at 13000 Distinguish between trisomy Downs syndrome and translocation Downs syndrome. (5 pt) Duplicate question Page 10 9. In the pea plant, the gene for purple flower color is dominant to its allele for red flower color. The gene for long pollen grain shape is dominant to its allele for round pollen grain shape. These genes are located on the same chromosome. In a two factor testcross experiment, a plant that was heterozygous for both genes was crossed with a plant having red flowers and round pollen grains. The following results were obtained. 445 Purple flower, long pollen 48 Purple flower, round pollen 52 Red flower, long pollen 455 Red flower, round pollen (a) In the heterozygous parent used to make the cross, was the linkage between the genes in the cis or the trans configuration? (2 pt) cis (b) What is the map distance between the genes for flower color and pollen shape? (4 pt) 10 map units (c) Is the distance that you calculated in part (a) most likely an overestimate or an underestimate of the actual map distance? Why? (4 pt) Underestimate, due to double crossovers 10. In corn, the gene for colored kernels (C) is dominant over its allele for colorless kernels (c). The gene for full kernels (Sh) is dominant over shrunken (sh). The gene for nonwaxy endosperm (Wx) is dominant over waxy (wx). These genes are located on the same chromosome. A plant heterozygous for all three traits was crossed with a triply homozygous recessive plant, and the following progeny were obtained. colored colorless colorless colored colorless colored colored colorless shrunken full shrunken full shrunken full shrunken full waxy waxy waxy waxy nonwaxy nonwaxy nonwaxy nonwaxy 305 128 18 74 66 22 112 275 (a) What is the correct order of these genes on the chromosome? (3 pt) Duplicate question (b) What are the linkages (cis or trans) between each pair of genes in the heterozygous parent? (3 pt) (c) What are the distances in map units between each pair of genes? Show your work. (4 pt) (d) What is the coefficient of coincidence and the interference? (4 pt) Page 11
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