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INSTANT DOWNLOAD COMPLETE TEST BANK WITH ANSWERS

 

 

Biotechnology 2nd Edition by David P. Clark  – Test Bank

 

 

Sample  Questions

 

Clark: Biotechnology, 2nd Edition

 

Chapter 3: Recombinant DNA Technology

 

 

  1. Escherichia coli is the workhorse in many molecular biology experiments. It is frequently necessary to obtain plasmid or chromosomal DNA from this organism. To do so:

*a. Lysozyme degrades the cell wall, detergent bursts the membranes, and finally phenol is used to remove protein.

  1. Lysozyme and RNAse are used to break open cells and release only the DNA.
  2. Phenol, lysozyme, and RNAse are added simultaneously prior to precipitation with ethanol.
  3. Phenol is required to precipitate the DNA.

 

  1. When agarose gel electrophoresis is used to study DNA,
  2. DNA is loaded near the positive electrode because it is negatively charged.
  3. Pieces of DNA are separated by size, with larger sizes moving further.
  4. Molecular weight standards are not needed as the movement of DNA in agarose is well understood.

*d. DNA moves from the negative electrode towards the positive electrode because it is negatively charged.

  1. Pulsed Field Gel Electrophoresis might be needed if the pieces are smaller than 10 kilobases.

 

  1. According to standard naming for restriction enzymes, the first enzyme isolated from Grampositivobacter skinius strain X would be called:
  2. Grs type I

*b. GrsX I

  1. GrsK X
  2. Grp SK

 

  1. Type I and Type II restriction enzymes differ in that
  2. Prokaryotes have type I and eukaryotes have type II.
  3. Type I produce blunt ends and Type II produce staggered ends.

*c. Type I enzymes cut 1000 base pairs away from the sequence they recognize whereas Type II cut within the recognition sequence.

  1. Type I enzymes cut only modified DNA but type II can cut any DNA.

 

  1. Restriction Fragment Length Polymorphisms:
  2. are identical among members of the same species
  3. are based on DNA fragment sizes observed after cutting with a DNA ligase.

*c. are based on differences seen when changes in the sequence fall in a restriction enzyme recognition site.

  1. all of the above are true.

 

  1. DNA probes used in a variety of experiments can be labeled for detection with all of the below EXCEPT:
  2. radioactive phosphate
  3. radioactive sulfur
  4. biotin
  5. digoxigenin

*e. all of the above can be used to label DNA

 

  1. The GC ratio of DNA is very important in many DNA technologies because:
  2. G and C will fluoresce whereas A and T will not.

*b. G and C will have 3 hydrogen bonds between them, resulting in a higher melting point.

  1. Restriction enzymes recognize sequences containing only G and C.
  2. The GC ratio is actually NOT important at all.

 

  1. In a Southern blot,
  2. RNA is transferred from a gel to a membrane the probed with single stranded probe.

*b. Hybrid DNA molecules are formed by transferring DNA from a gel to a membrane and probing with DNA.

  1. DNA or other molecular is spotted on a membrane and then hybridized with a probe.
  2. A probe is hybridized to DNA or RNA that is still in a cell.

 

  1. In a Northern blot,

*a. RNA is transferred from a gel to a membrane the probed with single stranded probe.

  1. Hybrid DNA molecules are formed by transferring DNA from a gel to a nylon membrane and probing with DNA.
  2. DNA or other molecular is spotted on a membrane and then hybridized with a probe.
  3. A probe is hybridized to DNA or RNA that is still in a cell.

 

  1. In a dot blot,
  2. RNA is transferred from a gel to a membrane the probed with single stranded probe.
  3. Hybrid DNA molecules are formed by transferring DNA from a gel to a nylon membrane and probing with DNA.

*c. DNA or other molecular is spotted on a membrane and then hybridized with a probe.

  1. A probe is hybridized to DNA or RNA that is still in a cell.

 

  1. In fluorescence in situ hybridization,
  2. RNA is transferred from a gel to a membrane the probed with single stranded probe.
  3. Hybrid DNA molecules are formed by transferring DNA from a gel to a nylon membrane and probing with DNA.
  4. DNA or other molecular is spotted on a membrane and then hybridized with a probe.

*d. A probe is hybridized to DNA or RNA that is still in a cell.

 

  1. A polylinker is a
  2. Method to determine if foreign DNA has been inserted into the lacZ gene.
  3. How many molecules of the vector can be found in a cell.
  4. Means to select for the cells that received vector.

*d. Region of DNA with many restriction enzyme sites.

 

  1. Copy number refers to the
  2. Method to determine if foreign DNA has been inserted into the lacZ gene.

*b. Approximate number of molecules of the vector that can be found in a single bacterial cell.

  1. Means to select for the cells that received vector.
  2. Region of DNA with many restriction enzyme sites.

 

  1. An antibiotic resistance gene is
  2. The reason people need to take antibiotics when they have a cold
  3. Found only in eukaryotes

*c. Is sometimes used to select for the bacterial cells that received vector during transformation

  1. A region of DNA with many restriction enzyme sites.

 

  1. Alpha complementation is a

*a. Method to determine if foreign DNA has been inserted into the lacZ gene.

  1. Method to determine if a gene is toxic to the host cell
  2. Region of DNA with many restriction enzyme sites.
  3. Match the following terms to useful traits for cloning vectors.

 

  1. A shuttle vector
  2. A series of vectors containing large pieces of DNA that can be used to sequence an entire genome.

*b. Contains origins of replication for two organisms and can be transferred back and forth between them.

  1. Viral genomes with many genes removed so that foreign DNA can be inserted packaged, and transferred to E. coli.
  2. Vector containing only viral cos sites plus foreign DNA.

 

  1. Gateway cloning vectors are

*a. A series of vectors that uses the integrase and excisionase from lambda phage to move genes from one vector to another.

  1. Contains origins of replication for two organisms and can be transferred back and forth between them.
  2. Viral genomes with many genes removed so that foreign DNA can be inserted packaged, and transferred to E. coli.
  3. Vector containing only viral cos sites plus foreign DNA.

 

  1. To construct a gene library, DNA can be inserted into coli by:
  2. Transformation
  3. Electroporation

*c. Both of the above.

  1. None of the above.

 

  1. A gene library can be obtained by
  2. Relying on plasmid incompatibility.

*b. Digesting (or partially digesting) purified DNA with a restriction enzyme, ligation of that DNA into a plasmid, and transformation of the mixture into E. coli.

  1. Screening E. coli for sequences similar to those of your organism.
  2. It is only possible in prokaryotes.

 

  1. Expression libraries differ from gene libraries in which of the following ways?
  2. There is no difference.
  3. Expression libraries contain sequences needed for both transcription and translation.
  4. Expression libraries can be based on cDNA.

*d. b. and c. are both true.

 

  1. Expression vectors:
  2. allow for the control of protein production when proteins might be toxic
  3. have promoters that can be turned on and off.
  4. may add protein tags to the protein to aid in purification

*d. all of the above are true.

  1. some of the above are true.

 

  1. Recombineering generates large vectors by using .
  2. integrase
  3. attB and attP sites

*c. lambda RED enzymes

  1. topoisomerase
  2. gateway technology

 

  1. What is the purpose of the entry vector in Gateway® cloning?
  2. creates attB and attP sites

*b. creates attL sites on both ends of the segment

  1. generates excisionase
  2. creates a polylinker
  3. toxin production

 

 

 

Clark: Biotechnology, 2nd Edition

 

Chapter 5: RNA-Based Technologies

 

 

  1. Antisense RNA is a
  2. RNA that can be found in the ribosome that can carry out catalysis.

*b. RNA with sequence complementary to an mRNA, this RNA will prevent translation.

  1. Small non-coding RNA molecules that modulate gene expression
  2. Small non-coding RNA that identifies an mRNA and triggers degradation

 

  1. Short-interfering RNA (siRNA) are
  2. RNAs that can be found in the ribosome that can carry out catalysis.
  3. RNAs with sequence complementary to an mRNA, this RNA will prevent translation.
  4. Small non-coding RNA molecules that modulate gene expression

*d. Small non-coding RNAs that identify an mRNA and trigger its degradation

 

  1. MicroRNAs are
  2. RNAs that can be found in the ribosome that can carry out catalysis.
  3. RNAs with sequence complementary to an mRNA, this RNA will prevent translation.

*c. Small non-coding RNA molecules that modulate gene expression

  1. Small non-coding RNAs that identify an mRNA and trigger its degradation

 

  1. Ribozymes are

*a. RNAs that can be found in the ribosome that can carry out catalysis.

  1. RNAs with sequence complementary to an mRNA, this RNA will prevent translation.
  2. Small non-coding RNA molecules that modulate gene expression
  3. Small non-coding RNAs that identify an mRNA and trigger its degradation

 

  1. RNAI and RNAII are sense and antisense partners controlling this process.
  2. Control of HIV gene expression
  3. Developmental control of nFGF

*c. Control of ColE1 replication

  1. Control of Neurospora circadian rhythm
  2. Control of X-chromosome inactivation

 

  1. Antisense mRNA binds to cellular mRNA during development of Xenopus laevis and degrades the message.
  2. Control of HIV gene expression

*b. Developmental control of nFGF

  1. Control of ColE1 replication
  2. Control of Neurospora circadian rhythm
  3. Control of X-chromosome inactivation

 

  1. Involves the use of xist
  2. Control of HIV gene expression
  3. Developmental control of nFGF
  4. Control of ColE1 replication
  5. Control of Neurospora circadian rhythm

*e. Control of X-chromosome inactivation

 

  1. Antisense env mRNA binds to the Rev response element, blocking production of protein.

*a. Control of HIV gene expression

  1. Developmental control of nFGF
  2. Control of ColE1 replication
  3. Control of Neurospora circadian rhythm
  4. Control of X-chromosome inactivation

 

  1. Regulated by antisense and sense mRNA for the frq
  2. Control of HIV gene expression
  3. Developmental control of nFGF
  4. Control of ColE1 replication

*d. Control of Neurospora circadian rhythm

  1. Control of X-chromosome inactivation
  2. Regulated by antisense and sense mRNA for the frq gene.

 

  1. Antisense oligonucleotides may alter aberrant splicing. Which two of the following are examples of human diseases and the gene involved with aberrant splicing.
  2. The env mRNA in HIV and the antisense oligonucleotides for splice site in beta-globin gene.

*b. The antisense oligonucleotide for Bcl-x splice sites in cancer and the antisense oligonucleotides for splice site in beta-globin gene.

  1. The antisense oligonucleotide for Bcl-x splice sites in cancer and the antisense oligonucleotide for the env MRNA in HIV.
  2. None of the above.

 

  1. Major challenges to studying the use of antisense technology in the lab include all of the following EXCEPT:
  2. Nonspecific interactions with other molecules in the cells.

*b. The sensitivity to of dsRNA to RNase H.

  1. Targeting regions of mRNA that may fold back on themselves.
  2. The decision to synthesize the oligonucleotides chemically or to clone antisense genes.

 

  1. Which of the following delivery methods are used to deliver antisense RNA to cells:
  2. Liposomes
  3. Cationic polymers
  4. Streptolysin
  5. Scrape loading

*e. All of the above

  1. Fill-in: With RNA interference (RNAi), ______ triggers _______ to degrade the mRNA into short interfering RNA or siRNA.
  2. siRNA, dicer
  3. antisense RNA, slicer

*c. dsRNA, dicer

  1. miRNA, drosha

 

  1. RNAi has been observed in many organisms and given many names prior to the elucidation of the mechanism. Which of the following terms has NOT been a name for RNAi?
  2. Quelling.

*b. Attenuation.

  1. Posttranscriptional Gene Silencing.
  2. Transcriptional Gene Silencing.
  3. All of the above ARE names for RNAi.

 

  1. MicroRNAs (miRNAs) are small RNAs that modulate gene expression by
  2. Cutting stem loops.
  3. Binding to target RNA and blocking translation.
  4. Blocking to the 3’UTR.
  5. All of the above.

*e. Some of the above.

 

  1. Methods are developed to deliver dsRNA into cells in order to study gene expression. In elegans, which of the following methods is used.
  2. Feeding them E. coli that is expressing the RNA
  3. Bathing C. elegans in RNA which they then take up into their bodies.
  4. Inject the RNA into the worms.

*d. All of the above work to deliver dsRNA to C. elegans.

  1. Some of the above will work to deliver RNA to C. elegans.

 

  1. Studying RNAi is more problematic in human cell culture than in Drosophila or elegans because:
  2. dsRNA cannot get into the cells
  3. Interferon is produced triggering RNA degradation.
  4. A potent antiviral response is triggered to the dsRNA
  5. All of the above are correct.

*e. Some of the above are correct.

 

  1. Ribozymes have been found to carry out which of the following processes.
  2. Removal of group I introns from other mRNA.
  3. Degrade RNA, especially the 5′ end of pre-tRNA
  4. Participate in the replication of viroids and satellite viruses.

*d. All of the above.

  1. Some of the above.
  2. Antisense combined with an engineered ribozyme would work in which of the following ways.
  3. Antisense RNA regions on the ends of the RNA find the target and then RNAse H cleaves it. The product is degraded by the ribozyme.
  4. Antisense RNA region in the middle of the RNA finds the target and then the ribozymes cleave it.

*c. Antisense RNA regions on the ends of the RNA find the target and then the ribozyme cleaves it.

  1. All of the above are mechanisms for ribozyme and antisense RNA function.
  2. None of the above are mechanisms for Ribozyme and antisense RNA function.

 

  1. Ribozymes can be evolved in vitro and have gained which of the following activities EXCEPT.
  2. Catalyzing ligations.
  3. Adding metal ions.
  4. Carry out nucleophilic attack at phosphoryl, carbonyl, and alkyl halides.
  5. Conversion of deoxyribonucleotides to ribonucleotides.

*e. All of the above are activities that evolved ribozymes have gained.

 

  1. Riboswitches work by alternating between different RNA secondary structures in an mRNA. Which of the following is NOT true of riboswitch examples
  2. Many are found in biosynthetic operons of bacteria

*b. With the thiamine riboswitch, the presence of thiamine is required for the production of mRNA.

  1. There are both attenuator mechanisms affecting transcription and translational mechanisms preventing translation off a full length mRNA
  2. Riboswitches can be affected by temperature (thermal stress).
  3. All of the above are true.

 

  1. Regulation of miRNA abundance in eukaryotes is aided with the help of .
  2. miRNA
  3. snoRNA
  4. gRNA
  5. crRNA

*e. circRNA

 

  1. ________ rescues stalled ribosomes in bacteria.
  2. Xist

*b. tmRNA

  1. piRNA
  2. snoRNA

 

  1. Which of the following RNAs are involved in splicing?
  2. TERC

*b. snRNA

  1. gRNA
  2. miRNA

 

  1. ____________ is responsible for transposon silencing in germ lines.
  2. tmRNA
  3. circRNA
  4. tRNA

*d. piRNA

 

  1. Dyskeratosis congenital results from a deletion in the protein or RNA portion of telomerase. The RNA portion of telomerase is called and functions                                     .
  2. TERT; to shorten the telomeres in aging cells

*b. TERC; as a template to increase the length of telomeres

  1. Xist; to inactive one X chromosome in mammalian females
  2. piRNA; to silence transposons in germ line cells
  3. lncRNA; to maintain genome stability

 

  1. All of the following statements about the CRISPR system is true except .
  2. The CRISPR system is not found in eukaryotes.
  3. CRISPR protects against viruses having either RNA or DNA genomes, hostile plasmids, and transposons.
  4. CRISPR is based upon memory of short sequences of nucleic acid that are stored on the bacterial chromosome.

*d. CRISPR targets specific sequences on double-stranded RNA.

  1. Stored sequences within the bacterial genome are transcribed and processed into crRNA that are then used by CAS nucleases as guides to seek and destroy foreign nucleic acid.

 

  1. Concerning PTEN expression, which non-coding RNA functions to convert the PTEN genome region into heterochromatin to prevent transcription?

*a. PTENpg1 antisense ?

  1. PTENpg1 antisense ?
  2. PTENpg1 sense
  3. DNMT3A
  4. EZH2

Clark: Biotechnology, 2nd Edition

 

Chapter 11: Protein Engineering

 

 

  1. Which of the following enzymes are used by industry and are potential targets for protein engineering:
    1. Invertase
    2. Rennet
    3. Catalase
    4. Amylase
    5. Glucose isomerase

*f. All of the above

  1. Some of the above

 

  1. Disulfide bonds are involved in which of the following properties of proteins
    1. Maintaining the 3D structure.
    2. Minimizing oxidative damage.
    3. Increasing the melting temperature of the protein

*d. All of the above.

  1. None of the above.

 

  1. In addition to disulfide bonds, other ways to increase the stability of a protein include:
    1. Replacing flexible amino acids like glycine with more rigid amino acids like proline
    2. Filling small cavities in the protein surface by replacing small hydrophobic amino acids with larger hydrophobic amino acids.
    3. Replacing asparagine or glutamine with aspartic acid or glutamic acid, respectively.

*d. All of the above.

  1. None of the above.

 

  1. With respect to the binding sites of lactate dehydrogenase, which of the following statement is true:
    1. The overall enzymatic activity is destroyed when any amino acid is changed.
    2. The binding of substrates can be changed but the binding of cofactors cannot be changed.
    3. The active site can be made larger and more hydrophobic but still only lactic acid can be the substrate.

*d. Changing a hydrophobic amino acid in the cofactor pocket to a positively charged amino acid will allow this enzyme to now prefer NADP rather than NAD.

 

  1. With respect to beta-galactosidase activity, which of the following statements is true:
    1. Beta-galactosidase is much smaller than most other hydrolytic enzymes.

*b. It may be possible to engineer a smaller version of beta-galactosidase that still functions.

  1. From an industrial viewpoint, it would be far better to manufacture a larger version of beta-galactosidase.
  2. All of these statements are true.

 

  1. Directed evolution is a technique used to later the function of enzymes without needing structural data. Which of the following approaches are used in directed evolution.
    1. Different domains of various proteins can be recombined together to create new enzymes.
    2. Target mutagenesis of the putative active site can be carried out to alter enzyme function.
    3. A cloned gene can be mutated using error-prone PCR and then new enzymes functions selected.

*d. All of the above are appropriate approaches.

  1. Some of the above are appropriate approaches.

 

  1. The steps involved in engineering new proteins that contain non-natural amino acids include
    1. Cloning and directed evolution of a gene for tRNA synthetase to create the new tRNA charged with the non-natural amino acid.
    2. Engineering genes for the new tRNA so it will bind to the amber codon.
    3. Cloning all of these evolved genes into Methanococcus jannaschii.
    4. All of the above.

*e. Some of the above.

 

  1. Which of the following alterations has been used to expand the genetic code?
  2. tRNAs have been developed that recognize four nucleotides rather than three
  3. tRNAs synthetases have been engineered to add non-natural amino acids to amber tRNAs
  4. “Photo-caged” amino acids replace natural amino acids during protein synthesis
  5. Non-natural amino acids are used to cross-link the engineered proteins

*e.  All of the above have been used

  1. Some of the above have been used

 

  1. Which of the following is not a role for non-natural amino acids in engineered proteins?
    1. Provide reactive chemical groups
    2. Provide reactive groups that are activated by UV light
    3. Provide a fluorescent group for visualizing the protein
    4. Provide a label for NMR spectroscopy

*e. Provide a new site for FokI endonuclease

 

  1. When the DNA binding domain of FokI is combined with the recognition domain of the Gal4 activator, the result is
    1. A recombinant protein with Zinc finger binding.
    2. A restriction enzyme that cuts right in the middle of the Gal4 DNA recognition sequence.

*c. A nuclease that recognizes the GAL4 DNA recognition sequence and cuts next to it.

  1. An endonuclease that cuts nonspecifically.

 

  1. Zinc finger domains are found in many regulatory proteins. Of the following properties, which is NOT a property of a zinc finger domain?
    1. Consists of 25-30 amino acids arranged around a zinc ion.
    2. Binding to cysteines and histidines holds the zinc ion in place.

*c. Each domain has nuclease activity

  1. One zinc finger motif binds 3 base pairs.

 

  1. DNA shuffling is a method of artificial evolution of new enzyme activities. Which of the following steps may not be needed?

*a. Mutating the gene and obtaining a mutant bank.

  1. Digesting to obtain different pieces of DNA
  2. Reassembling the pieces of DNA, generating novel combinations.
  3. Screening the new combinations for activity.

 

  1. DNA shuffling can also be performed using multiple related genes. Which of the following have been improved in this manner.
    1. Insulin
    2. Erythropoietin

*c. Beta-lactamase

  1. Zinc fingers

 

  1. Which of the following is NOT an example of a combinatorial protein library.
    1. DNA coding for various protein domains recombined in a variety of ways.
    2. Exons and alternate exons being recombined in a variety of ways.

*c. Random PCR products from all over a genome are recombined into pieces of DNA and cloned.

  1. PCR products corresponding to protein motifs are randomly combined.

 

  1. The basis of many biomaterials is protein engineering with elastin-like polypeptides. Which of the following is NOT a result of this engineering.
    1. The ability of a doctor to inject liquids that then form gels.

*b. Zinc finger domains are used to turn on elastin-like polypeptide genes.

  1. Peptides can promote cell migration and adherence.
  2. Movement and growth of vascular smooth muscle cells.

 

  1. Which of the following is NOT a domain associated with protein –protein binding
    1. Beta-sandwich
    2. Three-helix bundle

*c. Zinc finger motif

  1. Disulfide-bonded scaffolds
  2. Small scaffolds

 

  1. Which of the following non-natural amino acid is used for analysis of post-translational modifications?

*a. Serine phosphorylation

  1. Azidohomoalanine
  2. p-Benzoyl-L-phenylalanine
  3. Fluoro-amino acids

 

  1. Which of the following non-natural amino acids is used as an isotopic label in NMR spectroscopy?
    1. Serine phosphorylation
    2. Azidohomoalanine
    3. p-Benzoyl-L-phenylalanine

*d. Fluoro-amino acids

 

  1. Selenocysteine and pyrrolysine are genetically encoded amino acids

*a. True

  1. False

 

  1. Protein engineering may involve all of the following except .
  2. Random polypeptide libraries
  3. Orthogonal synthetase/tRNAs for the incorporation of non-natural amino acids
  4. Recombination of different protein domains
  5. De novo protein synthesis

*e. All of the above are methods of protein engineering.