Exam 12: Molecular Techniques and Biotechnology

Infection of potato crops by the fungus Phytophthora infestans is a constant menace to farmers. A biotechnological approach to prevent the recurrence of such an infestation is:

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Question 1

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The field of biotechnology changed little over the millennia until 1928, when Alexander Fleming discovered:

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Question 2

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Describe the whole-genome DNA-binding analysis using ChIP-on-chip technology.

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This technique is useful in determining all the in vivo binding sites for a protein in a given bacterial genome. It takes advantage of the existence of microarrays that contain amplified small DNA fragments that represent the entire genome of a bacterium. A putative DNA binding protein (for example, a transcription factor) will be specifically bound to certain sequences. Other proteins will bind as well. Proteins are cross-linked to DNA inside the cell. The DNA is extracted and sheared to small pieces. The DNA fragments bound to the protein of interest are immunoprecipitated with bead-bound antibodies to the protein and collected. Co-precipitated DNA is separated from the protein by heating. Linkers are added to both ends of the released DNA and PCR is used to amplify these DNA fragments with fluorescence-tagged primers designed to the linkers. The amplicons are hybridized to a microarray chip. Fluorescent spots on the chip will precisely indicate what genomic regions were originally bound to the protein and which genes it likely controls.

What kind of mutation is produced by transposable elements?

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Question 4

Traditionally, vaccines used to prevent microbial diseases have consisted of:

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Question 5

A major advantage of real-time PCR is that it can be used to quantify the original amount of RNA in a sample. What additional step must be accomplished in this particular application?

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Question 6

A protein of interest can be specifically detected on Western blots; this technique requires:

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Question 7

Nucleic acids are easily transferred from agarose gels onto a membrane using the physical principle of __________.

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Question 8

What is the principle of the electrophoretic mobility shift assay (EMSA)? What is its main application? Mention a possible limitation.

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Question 9

How are libraries of phage enzyme mutants generated?

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Question 10

In isolating acid-sensitive E. coli using tetracycline-resistant transposons, why is it necessary to screen first for tetracycline resistance and then for acid-sensitivity?

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Question 11

Microbes that grow at neutral pH but can survive at pH 2 for short periods are called:

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Question 12

How is bacterial DNA cross-linked to putative binding proteins for chromatin immunoprecipitation (ChIP)?

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Question 13

Why must secondary antibodies be elicited in an animal of a different species from that in which the primary antibodies were obtained?

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Question 14

You are planning to do some Northern blots and would like to use a nonradioactive label. You decide to use biotinylated DNA. What detection method would you use?

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Question 15

Describe advantages and disadvantages of using adenoviruses and adeno-associated viruses as vectors in gene therapy.

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Question 16

The phenomenon that allows a quencher dye to reduce the fluorescence of the reporter dye in real-time PCR techniques is called:

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Question 17

Which technique is used to select novel enzymes produced through directed evolution?

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Question 18

A screening of a GadA- $\beta$ -gal fusion protein in E. coli is performed on medium containing X-Gal. White and blue colonies are observed after incubation. Which of the following statements is NOT true regarding blue/white selection?

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Question 19

DNA fragments with specific sequences are detected by a technique known as __________.

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Question 20

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Exam 12: Molecular Techniques and Biotechnology

Infection of potato crops by the fungus Phytophthora infestans is a constant menace to farmers. A biotechnological approach to prevent the recurrence of such an infestation is:

The field of biotechnology changed little over the millennia until 1928, when Alexander Fleming discovered:

Describe the whole-genome DNA-binding analysis using ChIP-on-chip technology.

What kind of mutation is produced by transposable elements?

Traditionally, vaccines used to prevent microbial diseases have consisted of:

A major advantage of real-time PCR is that it can be used to quantify the original amount of RNA in a sample. What additional step must be accomplished in this particular application?

A protein of interest can be specifically detected on Western blots; this technique requires:

Nucleic acids are easily transferred from agarose gels onto a membrane using the physical principle of __________.

What is the principle of the electrophoretic mobility shift assay (EMSA)? What is its main application? Mention a possible limitation.

How are libraries of phage enzyme mutants generated?

In isolating acid-sensitive E. coli using tetracycline-resistant transposons, why is it necessary to screen first for tetracycline resistance and then for acid-sensitivity?

Microbes that grow at neutral pH but can survive at pH 2 for short periods are called:

How is bacterial DNA cross-linked to putative binding proteins for chromatin immunoprecipitation (ChIP)?

Why must secondary antibodies be elicited in an animal of a different species from that in which the primary antibodies were obtained?

You are planning to do some Northern blots and would like to use a nonradioactive label. You decide to use biotinylated DNA. What detection method would you use?

Describe advantages and disadvantages of using adenoviruses and adeno-associated viruses as vectors in gene therapy.

The phenomenon that allows a quencher dye to reduce the fluorescence of the reporter dye in real-time PCR techniques is called:

Which technique is used to select novel enzymes produced through directed evolution?

A screening of a GadA- β\betaβ -gal fusion protein in E. coli is performed on medium containing X-Gal. White and blue colonies are observed after incubation. Which of the following statements is NOT true regarding blue/white selection?

DNA fragments with specific sequences are detected by a technique known as __________.

Microbial Life: Origin and Discovery

Observing the Microbial Cell

Cell Structure and Function

Bacterial Culture, Growth, and Development

Environmental Influences and Control of Microbial Growth

Virus Structure and Function

Genomes and Chromosomes

Transcription, Translation, and Bioinformatics

Gene Transfer, Mutations, and Genome Evolution

Molecular Regulation

Viral Molecular Biology

Energetics and Catabolism

Respiration, Lithotrophy, and Photolysis

Biosynthesis

Food and Industrial Microbiology

Origins and Evolution

Bacterial Diversity

Archaeal Diversity

Eukaryotic Diversity

Microbial Ecology

Microbes and the Global Environment

Human Microbiota and Nonspecific Host Defenses

The Adaptive Immune Response

Microbial Pathogenesis

Microbial Diseases

Antimicrobial Chemotherapy

Clinical Microbiology and Epidemiology

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A screening of a GadA- $\beta$ -gal fusion protein in E. coli is performed on medium containing X-Gal. White and blue colonies are observed after incubation. Which of the following statements is NOT true regarding blue/white selection?