Exam 12: Recombinant Dna, Cloning, Chimeric Genes, and Synthetic Biology

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Reporter genes such as those for green fluorescent protein (GFP) are found on many commercial plasmids. These genes are useful to determine:

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Question 1

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All are true for cDNA libraries EXCEPT:

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Question 2

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Explain how the CRISPR/Cas9 system can be used to edit the genome.

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Question 3

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A nucleotide sequence in any genome can be edited with an engineered version of the CRISPR/Cas9 system found in the bacterium Streptococcus pyogenes. S. pyogenes Cas9 is an RNA-guided endonuclease that catalyzes cleavage of both strands of a dsDNA at a site determined by base-pairing between the Cas9-associated guide RNA (gRNA) and the target DNA nucleotide sequence. Since a gRNA of any desired nucleotide sequence is easily made, any DNA sequence is a legitimate target for Cas9 endonucleolytic cleavage. DNA repair systems such as nonhomologous end joining (NHEJ) and homology-directed repair (HDR) will repair the double-stranded breaks, and the genes can be screened for the hoped-for changes in nucleotide sequence.

Looking at all of the genes that are activated during a major metabolic shift or during embryogenesis and development of organisms is called:

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Question 4

The correct sequence for colony hybridization experiments is: A.A replica of the bacterial colonies is obtained on an absorbent disC. B.Autoradiography of the disc reveals probe complementary DNA. C.Host bacteria with plasmid are plated and allowed to grow overnight. D.The disc is treated with alkali. E.The disc is reacted with labeled probe.

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Question 5

If DNA fragments of about 4 kb are to be cloned, which vector would be most useful?

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Question 6

The original development of PCR did not use the Taq polymerase. What is the reason that the Taq polymerase is currently used?

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Question 7

All are strategies for human gene therapy EXCEPT:

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Question 8

One of the problems with plasmids like pUR278 is that there is no true marker for determining whether or not the plasmid contains the inserted gene since the cloning site does not lie within any specific gene. Which of the following would be an extremely useful tool for rapid selection of those bacteria that have taken up a plasmid with the desired DNA insert?

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Question 9

The advantages of using the pET plasmid system include ____.

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Question 10

In a DNA microarray, how many different oligonucleotides can be produced if each oligonucleotide is 20 nucleotides long?

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Question 11

If the pBR322 were treated with PstI in order to insert a gene into the plasmid, which of the following would be correct regarding cells that were transformed with the resulting vector?

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Question 12

The most promising vector for human gene therapy is ____.

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Question 13

All are characteristic of plasmids EXCEPT:

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Question 14

Discuss polymerase chain reaction-based (PCR-based) mutagenesis.

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Question 15

Shuttle vectors have the property that they:

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Question 16

RT-PCR differs from basic PCR in that:

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Question 17

Hybrid proteins or fusion proteins are produced by:

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Question 18

How are chimeric plasmids constructed?

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Question 19

The correct sequence of procedures in the Southern blotting (hybridization) technique is: A.hybridization with radioactive probE. B.agarose gel electrophoresis and visualize bands. C.transfer (blot) to nitrocellulose filter. D.digest DNA with restriction nucleases. E.expose filter to X-ray film, develop and observe.

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Question 20

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Exam 12: Recombinant Dna, Cloning, Chimeric Genes, and Synthetic Biology

Reporter genes such as those for green fluorescent protein (GFP) are found on many commercial plasmids. These genes are useful to determine:

All are true for cDNA libraries EXCEPT:

Explain how the CRISPR/Cas9 system can be used to edit the genome.​

Looking at all of the genes that are activated during a major metabolic shift or during embryogenesis and development of organisms is called:

If DNA fragments of about 4 kb are to be cloned, which vector would be most useful?

The original development of PCR did not use the Taq polymerase. What is the reason that the Taq polymerase is currently used?

All are strategies for human gene therapy EXCEPT:

The advantages of using the pET plasmid system include ____.

In a DNA microarray, how many different oligonucleotides can be produced if each oligonucleotide is 20 nucleotides long?

If the pBR322 were treated with PstI in order to insert a gene into the plasmid, which of the following would be correct regarding cells that were transformed with the resulting vector?

The most promising vector for human gene therapy is ____.

All are characteristic of plasmids EXCEPT:

Discuss polymerase chain reaction-based (PCR-based) mutagenesis.​

Shuttle vectors have the property that they:

RT-PCR differs from basic PCR in that:

Hybrid proteins or fusion proteins are produced by:

How are chimeric plasmids constructed?​

The Facts of Life: Chemistry Is the Logic of Biological Phenomena

Water: the Medium of Life

Thermodynamics of Biological Systems

Amino Acids and the Peptide Bond

Proteins: Their Primary Structure and Biological Functions

Proteins: Secondary, Tertiary, and Quaternary Structure

Carbohydrates and the Glycoconjugates of Cell Surfaces

Lipids

Membranes and Membrane Transport

Nucleotides and Nucleic Acids

Structure of Nucleic Acids

Enzymeskinetics and Specificity

Mechanisms of Enzyme Action

Enzyme Regulation

Molecular Motors

Metabolism: an Overview

Glycolysis

The Tricarboxylic Acid Cycle

Electron Transport and Oxidative Phosphorylation

Photosynthesis

Gluconeogenesis, Glycogen Metabolism, and the Pentose Phosphate Pathway

Fatty Acid Catabolism

Lipid Biosynthesis

Nitrogen Acquisition and Amino Acid Metabolism

Synthesis and Degradation of Nucleotides

Metabolic Integration and Organ Specialization

Dna Metabolism: Replication, Recombination, and Repair

Transcription and the Regulation of Gene Expression

Protein Synthesis

Completing the Protein Life Cycle: Folding, Processing, and Degradation

The Reception and Transmission of Extracellular Information

Filters

Explain how the CRISPR/Cas9 system can be used to edit the genome.

Discuss polymerase chain reaction-based (PCR-based) mutagenesis.

How are chimeric plasmids constructed?