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Exam 19: Molecular Genetic Analysis and Biotechnology

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Question 21

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You are trying to determine whether a certain RFLP marker is linked to a specific disease gene in dogs. A cross between a diseased male and a healthy female produces nine offspring. You isolate genomic DNA from parents and offspring, digest with EcoRI, radiolabel a portion of RFLP marker to use as a probe, and perform a Southern analysis. The pedigree and autoradiogram results are shown in the following figure. Assume that the disease is autosomal dominant. (a) Explain the relationship between the crossing results (i.e., diseased versus healthy) and the RFLP patterns.

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Question 22

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Cronin et al. (Science, 2009, 325: 340-343) used RNA interference to study the immune response of the fruit fly, Drosophila melanogaster, to the bacterial pathogen Serratia marcescens. They identified several members of the JAK-STAT cell-cell signaling pathway in their analysis. a) The PIAS gene is a negative regulator of JAK-STAT signaling (that is, it inhibits the signaling pathway). RNAi to block PIAS activity causes flies to die significantly earlier than control flies when exposed to bacteria. Does JAK-STAT signaling appear to protect the flies from the bacteria? Explain. b) The upd gene encodes a ligand that activates JAK-STAT signaling. RNAi that blocks upd function causes flies to survive longer than control flies when exposed to bacteria. Explain how this is or is not consistent with your answer from part (a).

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a) Yes, JAK-STAT signaling appears to pr...

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Question 23

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Use the following to answer questions Figure A below shows a restriction map of a rare prokaryotic gene with its direction of transcription indicated by the arrow. Figure B shows the unique restriction sites contained within a plasmid-cloning vector. The blackened region in Figure A represents the amino acid coding sequence of a protein that can be used in humans as a vaccine. The striped region in Figure B is a highly active, constitutive (unregulated) prokaryotic promoter region. Letters indicate the cleavage sites for different restriction enzymes. Known DNA sequences are indicated by short thick lines. -After trying to isolate and then insert the coding region (Figure A) under the control of the indicated promoter in the cloning vector (Figure B), you found that all the transformed bacterial cells contained either one of two smaller portions of the coding region for the gene, and some of the fragments were inserted backward (with regard to reading frame) into the cloning vector. How would you explain these observations?

Use the following to answer questions A fragment of DNA is cloned into a plasmid with a sequencing primer-binding site. After dideoxy sequencing, the gel pattern shown in this diagram is obtained. -On the strand that acted as the template in the sequencing reaction, what base of the cloned fragment was closest to the primer?

Which of the following is a current concern of genetic testing? (Select all that apply.)

What is the purpose of Taq polymerase in a PCR reaction?

Use the following to answer questions A fragment of DNA is cloned into a plasmid with a sequencing primer-binding site. After dideoxy sequencing, the gel pattern shown in this diagram is obtained. -What was the sequence of the DNA strand that acted as the template in the sequencing reaction?

Which of the following statements does NOT describe a challenge of working at the molecular level?

Antibodies are to Western blots as _____ is/are to Southern blots.

Which of the following would be MOST appropriate for cloning a gene that is 300 kb in size?

Which of the following statements is NOT correct regarding type II restriction enzymes?

A scientist is studying a normal tissue sample and a cancerous tissue sample. What method might she use to determine whether the transcription of gene X is upregulated in the cancerous tissue sample?

The haploid human genome contains about 3 × 10⁹ nucleotides. On average, how many DNA fragments would be produced if this DNA was digested with restriction enzyme PstI (a 6-base cutter)? RsaI (a 4-base cutter)? How often would an 8-base cutter cleave?

Which of the following represents an appropriate cloning vector for cloning a gene into a bacterial cell?

Explain what an RFLP is. Why do RFLPs behave like "codominant" markers?

Which of the following traits of type I restriction enzymes make them unsuitable for recombinant DNA technology? (Select all that apply.)

One technique for finding a gene of interest involves first generating a genetic map to find the general location of the gene and then identifying the specific location of the gene. What is this technique called?

Use the following to answer questions A scientist carries out a ligation reaction designed to insert a foreign piece of DNA into a plasmid that contains the front end of the lacZ gene within the multiple cloning site (MCS) . He carries out three transformations in parallel with lacZ^- bacteria. The three transformations (i, ii, and iii) contain the following: i. sterile water ii. a sample of the original plasmid iii. the ligation reaction He then plates the transformations on medium containing X-gal and ampicillin. -The scientist observed colonies on plate (i) . Which of the following might be reasons(s) for this? (Select all that apply.)

You are handling a paternity lawsuit brought against five potential fathers by a woman. You isolated DNA from the mother, the child, and all the potential fathers. After using PCR to amplify specific polymorphic loci from each individual, you run a gel of the amplified products and stain with ethidium bromide to visualize the DNA fingerprints (shown below). Mo = mother; Ch = child; M1-M5 = potential fathers. B1-B10 indicate marker bands present in the child. Do these results suggest that any of the men could be the child's biological father? Explain your answer.

Use the following to answer questions Figure A below shows a restriction map of a rare prokaryotic gene with its direction of transcription indicated by the arrow. Figure B shows the unique restriction sites contained within a plasmid-cloning vector. The blackened region in Figure A represents the amino acid coding sequence of a protein that can be used in humans as a vaccine. The striped region in Figure B is a highly active, constitutive (unregulated) prokaryotic promoter region. Letters indicate the cleavage sites for different restriction enzymes. Known DNA sequences are indicated by short thick lines. -Explain how you would isolate and then insert the coding region (Figure A) under the control of the indicated promoter in the cloning vector (Figure B) to produce large amounts of the protein in bacterial cells. Assume that the cloning vector carries the gene for tetracycline (an antibiotic) resistance.