mcb3020 4c 1, mcb3020 4c 2
Which of the following temperatures would be appropriate for the annealing step of PCR? Multiple choice question. 95OC 72OC 50OC 27OC
50OC
Which of the following temperatures would be appropriate for the extension/synthesis step of PCR? Multiple choice question. 95OC 72OC 50OC 27OC
72OC
What happens in the extension step of PCR? Multiple choice question. Sticky ends of complementary template fragments are covalently joined by DNA ligase. The hydrogen bonds holding the two strands of DNA are broken to produce single-stranded DNA. Primers hydrogen bond with their complementary sequences in the template DNA. A thermostable DNA polymerase adds dNTPs to the 3'-OH end of the primers.
A thermostable DNA polymerase adds dNTPs to the 3'-OH end of the primers.
Which of the following is NOT a component of a PCR reaction mixture? Multiple choice question. Oligonucleotide primers Taq polymerase ATP dNTPs
ATP
Bacterial or yeast ___ ____ are cloning vectors that are useful for cloning very large amounts of DNA such as the amount necessary to build a genomic library.
Artificial chromosomes
Which type of cloning vector can contain the largest amount of insert DNA? Multiple choice question. Artificial chromosomes Cosmids Phage vectors Plasmids
Artificial chromosomes
Which two of the following are advantages of plasmid cloning vectors? Multiple select question. Can accommodate up to 1,000 kb insert size Can be packaged into lambda phage particles Replicate once per cell cycle Autonomous replication Easy to purify
Autonomous replication Easy to purify
Cas9 genome editing is also referred to as _____. Multiple choice question. CRISPR real-time editing cloning seamless cloning
CRISPR
Genome editing techniques have been developed to exploit the CRISPR system of bacteria and archaea. Only the _____ component of the system is used in editing. Multiple choice question. DNA repair DNA:RNA hybrid CRISPR Cas9
Cas9
Cas9 endonuclease differs from restriction endonucleases in that _____. Multiple choice question. Cas9 probes DNA sequence by amino acid interaction Cas9 identifies a cut site by hybridizing a guide RNA Cas9 cuts both strands of DNA Cas9 recognizes a 4-8 base pair sequence of DNA
Cas9 identifies a cut site by hybridizing a guide RNA
In a microbial cell, invading DNA from bacteriophage or plasmids is targeted for destruction by ___. Multiple choice question. Cas9 nuclease RecA phage integrases topoisomerase
Cas9 nuclease
A(n) --- protein is a polypeptide that consists of two parts, each from a different source.
Chimeric
Which of the following would be useful to a scientist who wanted to put DNA from one organism into a different organism and then make many copies of it? Multiple choice question. Mechanical vector Support vector Cloning vector Dispersal vector
Cloning vector
Which two of the following are characteristics of phage vectors? Multiple select question. Can hold up to 1,000 kb of insert DNA Contains unique restriction sites Contains phage genes for host lysis Contains plasmid ori Contains yeast telomere sequences
Contains unique restriction sites Contains phage genes for host lysis
Which of the following is a hybrid cloning vector, containing DNA sequences from both plasmids and phages? Multiple choice question. YAC Cosmid YEp24 BAC
Cosmid
Which enzyme forms phosphodiester bonds between cloned DNA fragments and the cloning vector? Multiple choice question. DNA polymerase β-lactamase restriction endonuclease DNA ligase
DNA ligase
Which three of the following properties apply to green fluorescent protein? Multiple select question. Easily cloned and expressed in any organism Encoded by a single gene From the fluorescent mushroom Mycena chlorophos Encoded in a two gene operon Easily cloned but can only fluoresce in bacteria From the jellyfish Aequorea victoria
Easily cloned and expressed in any organism Encoded by a single gene From the jellyfish Aequorea victoria
Which of the following are steps of the polymerase chain reaction? Multiple select question. Extension Annealing Denaturation Digestion Transformation Ligation
Extension Annealing Denaturation
Which statement about polyhistidine-tagged protein purification is true? Multiple choice question. Histidine residues preferentially bind to solid phase materials with exposed chlorine or phosphate atoms Histidine residues preferentially bind to solid phase materials with exposed cobalt or nickel atoms Histidine residues preferentially bind to solid phase materials with exposed aluminum or thallium atoms
Histidine residues preferentially bind to solid phase materials with exposed cobalt or nickel atoms
Which three of the following are components of a cosmid? Multiple select question. Multicloning site cos site Telomere Origin of transfer Selectable marker
Multicloning site cos site Selectable marker
Which three of the following are important features shared by all types of cloning vectors? Multiple select question. Origin of transfer Origin of replication 100 - 1000 kb insert size Multicloning site URA3 gene Selectable marker
Origin of replication Multicloning site Selectable marker
Which four of the following are major types of cloning vectors? Multiple select question. Plasmids Introns Artificial chromosomes Cosmids Operons Bacteriophages Mobile genetic elements
Plasmids Artificial chromosomes Cosmids Bacteriophages
Which technique can amplify a specific sequence of DNA from a complex mixture of DNA sequences? Multiple choice question. Oligonucleotide sequencing Polymerase chain reaction Gel electrophoresis Reverse transcription
Polymerase chain reaction
What happens in the annealing step of PCR? Multiple choice question. The hydrogen bonds holding the two strands of DNA are broken to produce single-stranded template DNA. Radioactive probes bind to their target sequences. Primers hydrogen bond with their complementary sequences in the template DNA. A thermostable DNA polymerase adds dNTPs to the 3'-OH end of the primers.
Primers hydrogen bond with their complementary sequences in the template DNA.
Oligonucleotide sequences complementary to sequences flanking the DNA sequences to be amplified in the polymerase chain reaction are called ___
Primers or DNA primers
Which two of the following statements about protein purification by polyhistidine tagging are true? Multiple select question. Proteins with multiple histidine residues will bind to a solid matrix containing exposed nickel or cobalt atoms Proteins with multiple histidine residues will bind to a solid matrix containing exposed chlorine or fluorine atoms Histidine has a high affinity for metal ions Histidine has a low affinity for metal ions
Proteins with multiple histidine residues will bind to a solid matrix containing exposed nickel or cobalt atoms Histidine has a high affinity for metal ions
Which two statements describe a selectable marker? Multiple select question. Provides a mechanism for discriminating between transformed and non-transformed host cells Provides multiple unique restriction sites for inserting DNA fragments into the cloning vector Maintains selective pressure for the presence of the cloning vector in the host cell Allows for the replication of the cloning vector independent of the host cell chromosome
Provides a mechanism for discriminating between transformed and non-transformed host cells Maintains selective pressure for the presence of the cloning vector in the host cell
What is a plasmid multicloning site? Multiple choice question. Provides multiple restriction sites for inserting DNA fragments into the cloning vector. Provides a mechanism for discriminating between transformed and non-transformed host cells. Maintains selective pressure for the presence of the cloning vector in the host cell. Allows for the replication of the cloning vector independent of the host cell chromosome.
Provides multiple restriction sites for inserting DNA fragments into the cloning vector.
Restriction endonucleases (REs) differ from Cas9 endonuclease in that _____. Multiple choice question. REs remove the nitrogenous base from the DNA REs cut only one strand of DNA REs identify a cut site by hybridizing a guide RNA REs recognize a 4-8 base pair palindromic DNA sequence
REs recognize a 4-8 base pair palindromic DNA sequence
The most commonly used thermostable polymerase in the polymerase chain reaction is ____ polymerase.
Taq
The most commonly used thermostable DNA polymerase for the polymerase chain reaction is ____. Multiple choice question. Taq polymerase EcoRI polymerase RNA primase reverse transcriptase
Taq polymerase
Which of the following is NOT an important feature of all types of cloning vectors? Multiple choice question. Origin of replication Selectable marker Region with unique restriction sites Telomere sequence at each end
Telomere sequence at each end
What happens in the denaturation step of PCR? Multiple choice question. A thermostable DNA polymerase adds dNTPs to the 3'-OH end of each annealed primer. Sticky ends of complementary template fragments are covalently joined by DNA ligase. The hydrogen bonds holding the two strands of DNA are broken to produce single-stranded template DNA. Primers hydrogen bond with their complementary sequences in the template DNA.
The hydrogen bonds holding the two strands of DNA are broken to produce single-stranded template DNA.
Which of the following is NOT a major type of cloning vector? Multiple choice question. Plasmid Cosmid Bacteriophage Artificial chromosome Yeast plastid
Yeast plastid
In Cas9 genome editing, a guide RNA is ___. Multiple choice question. a component of Cas9 nuclease that cuts DNA at the CRISPR locus a component of Cas9 nuclease that specifies where to cut DNA a 20 base RNA that binds transcripts to interfere with translation a 20 base RNA that directs ribosomes to translate a CRISPR locus
a component of Cas9 nuclease that specifies where to cut DNA
His-tagging is a protein purification method that involves _________. Multiple choice question. using an RNA polymerase that uses a polyhistidine coding sequence as a strong promoter site using a expression vector that requires histidine as the inducer molecule adding a series of histidine residues to one end of the protein fusing a chemical group to a histidine side chain in the protein
adding a series of histidine residues to one end of the protein
Adding six histidine residues to a protein involves _____. Multiple choice question. modifying the ribosome to promote histidine incorporation adding six codons to the gene encoding the protein adding six nucleotides to the gene encoding the protein adding a new tRNA synthetase to the host cell
adding six codons to the gene encoding the protein
When selecting bacterial cells that have taken up a recombinant DNA molecule created in pUC19, the antibiotic ampicillin is included in the medium to ensure that _____. Multiple choice question. all colonies carry a plasmid with an inserted DNA fragment all colonies carry the plasmid contamination is minimized ampicilin resistance will not be transferred
all colonies carry the plasmid
The selectable marker for the pUC19 plasmid is _____. Multiple choice question. tetR lacZ URA3 ampR
ampR
In the plasmid pUC19, the selectable marker is a gene that encodes resistance to the antibiotic .
ampicillin (amp^R)
Making a large number of copies of a specific DNA sequence is referred to as ____. Multiple choice question. amplification reverse transcription oligonucleotide synthesis sequencing
amplification
Making a large number of copies of a specific DNA sequence is referred to as ____. Multiple choice question. amplification sequencing reverse transcription oligonucleotide synthesis
amplification
The process to produce a large number of copies of a specific DNA sequence is referred to as DNA ___
amplification
During PCR, the temperature is maintained at about 50°C in one step of each cycle to _____. Multiple choice question. denature the DNA template anneal the primers synthesize a new DNA strand
anneal the primers
In the second step of PCR, called ____, the oligonucleotide primers base pair with their complementary sequences in the single-stranded template DNA.
annealing
CRISPR loci are found in ___ and ___. Multiple select question. viruses animals archaea protozoans bacteria
archaea bacteria
Bacterial colonies transformed by pUC19 that lack a DNA insert, will have a color of ____ when grown in the presence of X-gal.
blue
A hybrid polypeptide, composed of portions of two or more proteins from different sources, is referred to as a(n) ______ protein. Multiple choice question. homodimeric heterodimeric anabolic chimeric
chimeric
A DNA molecule, such as a plasmid, that carries recombinant DNA into a host cell for replication is referred to as a(n) __vector
cloning
Plasmids can be introduced into bacterial cells by ___ and ___. Multiple select question. transduction conjugation transformation infection transposition
conjugation transformation
Seamless cloning refers to _____. Multiple choice question. the use of PCR to generate DNA fragments for cloning the use of several enzymes concurrently to act on a DNA molecule in vitro propagating a piece of DNA in a cloning vector to make many copies of it creating a recombinant DNA molecule with no scars from restriction sites
creating a recombinant DNA molecule with no scars from restriction sites
Which four of the following are components of a PCR reaction mixture? Multiple select question. dNTPs Template DNA Taq polymerase ATP Reverse transcriptase Primers restriction endonuclease
dNTPs Template DNA Taq polymerase Primers
In the first step of PCR, called ___, the double-stranded template DNA is separated into single strands by a temperature increase to 95°C.
denaturation
Place the steps of the polymerase chain reaction in the correct order. annealing denaturation extension
denaturation annealing extension
When screening bacterial cells that have taken up a recombinant DNA molecule created in pUC19, the chemical X-gal is included in the medium to _____. Multiple choice question. ensure that all colonies carry the plasmid ensure that all colonies carry a plasmid with an inserted DNA fragment distinguish colonies whose plasmids have an insert from those that don't have the insert distinguish colonies that can metabolize lactose from those that can't
distinguish colonies whose plasmids have an insert from those that don't have the insert
Two commonly used methods for introducing recombinant DNA into host organisms are ___ and ___. Multiple select question. electroporation chemical transformation cell fusion conjugation electrophoresis
electroporation chemical transformation
CRISPR/Cas systems are particularly useful for editing eukaryotic genomes because _____. Multiple choice question. eukaryotic DNA is readily accessible within the nucleus eukaryotic genomes lack CRISPR/Cas systems CRISPR/Cas systems work most effectively at a telomere almost no eukaryotic DNA originates from viruses
eukaryotic genomes lack CRISPR/Cas systems
In the third step of PCR, called ___, Taq polymerase adds dNTPs to the 3'-OH end of the primers.
extension
The multicloning site in pUC19 is located in the 5' region of a gene called β-___, which encodes the enzyme that cleaves lactose into glucose and galactose. (Leave out any Greek symbols.)
galactosidase, lacZ, or gal
The modification of the genetic code of a living organism is called _____. Multiple choice question. DNA cloning mutagenesis genomic sequencing genetic engineering
genetic engineering
A(n) ___ library contains the genetic information from a single organism.
genomic
A collection of recombinant vectors that contains fragments of one organism's entire genome is called a(n) ______. Multiple choice question. genomic library cDNA library metagenomic library expression library
genomic library Reason: A metagenomic library derives from the environment and is very likely to have genomic fragments from multiple organisms.
A protein originally from jellyfish that can be used to study promoter activity or protein localization is the ___ ___ protein.
green fluorescent
CRISPRs are _____. Multiple choice question. enzymes that hybridize to a specific segment of DNA groups of short palindromic repetitive sequences DNA:RNA hybrid molecules that form during viral infection viral DNA sequences that have previously infected a cell
groups of short palindromic repetitive sequences
Cas9 consists of a polypeptide and a _____. Multiple choice question. guide RNA CRISPR Mg+2 ion palindromic repeat
guide RNA
To edit a eukaryotic genome by CRISPR/Cas, two components must be added to the cell, ___ and ___. Multiple select question. guide RNA (gRNA) Cas9 holoenzyme Cas9 apoenzyme CRISPR repeats rRNA
guide RNA (gRNA) Cas9 apoenzyme
A gene from one organism that has been transferred to a different organism is referred to as a _____. Multiple choice question. hemizygous gene heterologous gene homologous gene heterozygous gene
heterologous gene
A(n) ___ gene is a gene from one organism that has been cloned into another organism.
heterologous or recombinant
One strategy for purifying an expressed protein from a mixture of bacterial proteins is to modify the protein's gene to encode a series of ___ amino acid residues at the N- or C-terminus of the protein.
histidine
Sticky ends that have been created by the same restriction endonuclease can associate with each other by _____. Multiple choice question. covalent bonds hydrogen bonds Van der Waals interactions ionic bonds
hydrogen bonds
Sticky ends that have been created by the same restriction endonuclease can associate with each other by _____. Multiple choice question. hydrogen bonds covalent bonds ionic bonds Van der Waals interactions
hydrogen bonds
In bacterial and archaeal chromosomes, the guide RNA is encoded _____. Multiple choice question. near the origin of replication at random locations in the ribosomal RNA operon in the CRISPR locus
in the CRISPR locus
The origin of replication allows the plasmid to replicate _____. Multiple choice question. independently of the host chromosome only when an inducer is present only once per cell generation
independently of the host chromosome
In pUC19, which gene sequence is interrupted by the presence of insert DNA? Multiple choice question. ori lacZ ampR URA3
lacZ
Once the sticky ends of the cloned DNA and cloning vector have formed complementary base pairs by hydrogen bonding, the two fragments can be covalently joined by adding the enzyme DNA ___ to the reaction mixture.
ligase
Sequences in the CRISPR locus of a bacterial chromosome derive primarily from _____. Multiple choice question. anabolic operons transfer RNAs mobile genetic elements the Cas9 nuclease
mobile genetic elements
Genetic engineering includes _____. [Choose three.] Multiple select question. treating a patient with an antiviral drug modifying bacteria to produce vitamins vaccinating livestock against infectious disease crossing crop plants to improve yield breeding animals for desirable traits purifying antibiotics from organisms that produce them
modifying bacteria to produce vitamins crossing crop plants to improve yield breeding animals for desirable traits
The region of a cloning vector that contains several restriction sites intended for the insertion of a DNA sequence is called the ___ site.
multicloning,
The ___ of ___ allows a plasmid to replicate independently of the host chromosome.
origin replication
Engineered genomes of bacterial viruses that contain unique restriction sites and the genes necessary for host lysis are called ___ vectors.
phage
Small circular double-stranded DNA cloning vectors that replicate autonomously and are easy to purify are
plasmids
The ___ ____ reaction is a technique that allows the production of a large number of copies of a specific DNA sequence from a complex mixture of sequences.
polymerase chain
Oligonucleotides that serve as the 3'-OH needed for DNA synthesis in the polymerase chain reaction are called _____. Multiple choice question. initiators primers promoters enhancers
primers
The normal cellular function of the Cas9 nuclease is to _____. Multiple choice question. assist in homologous recombination adjust cellular supercoiling protect the cell from viruses eliminate antibiotic resistance
protect the cell from viruses
An enzyme that recognizes and cleaves specific nucleotide sequences is called a(n) ____. Multiple choice question. DNA glycosylase topoisomerase DNA ligase restriction endonuclease
restriction endonuclease
Plasmid construction methods that use PCR instead of restriction endonucleases to specify the desired junctions between DNA molecules are termed _____. Multiple choice question. phenotypic rescue artificial chromosomes seamless cloning expression vectors
seamless cloning
A gene whose presence confers the ability to survive in particular conditions is called a(n) ______. Multiple choice question. polylinker selectable marker multicloning site origin of replication
selectable marker
A gene whose presence confers the ability to survive in particular conditions is called a(n) ______. Multiple choice question. selectable marker origin of replication polylinker multicloning site
selectable marker
Plasmids and other vectors that contain two origins of replication recognized by different host organisms are called ___ vectors.
shuttle
A plasmid that contains two origins of replication recognized by different host organisms is called a _____. Multiple choice question. expression vector replicon shuttle vector cloning vector
shuttle vector
During PCR, the temperature is maintained at about 68-72°C in one step of each cycle to _____. Multiple choice question. denature the DNA template anneal the primers synthesize a new DNA strand
synthesize a new DNA strand
The extreme specificity that is key to Cas9 genome editing is conferred by ___. Multiple choice question. the donor DNA the guide RNA the Cas9 nuclease the genomic DNA
the guide RNA
Plasmid copy number refers to _____. Multiple choice question. the number of plasmid molecules present in a cell the number of genes in a plasmid the length of DNA insert that a plasmid can tolerate the fraction of a population that has taken up a plasmid
the number of plasmid molecules present in a cell
Conjugation, transformation, and electroporation are all methods for _____. Multiple choice question. separating DNA fragments according to size combining DNA fragments into a recombinant DNA molecule transferring a plasmid cloning vector to a microbe plasmid cloning vector replication in a new microbe
transferring a plasmid cloning vector to a microbe
Conjugation, transformation, and electroporation are all methods for _____. Multiple choice question. separating DNA fragments according to size plasmid cloning vector replication in a new microbe combining DNA fragments into a recombinant DNA molecule transferring a plasmid cloning vector to a microbe
transferring a plasmid cloning vector to a microbe
Chemical ___ and ___ (a technique that uses high-voltage electricity), are two commonly employed methods for the introduction of recombinant DNA into host organisms.
transformation Blank 2: electroporation
True or false: Sticky ends created by the same restriction enzyme can hydrogen bond with each other. True false question. True False
true
Adding the codons for six histidines to a gene can be performed in two ways, Multiple select question. using an oligonucleotide primer to direct the Cas9 nuclease to the site of histidine codon placement using a transpeptidase enzyme and a high concentration of histidine using an expression vector that has the six codons adjacent to a multiple cloning site using a restriction enzyme whose recognition sequence includes histidine codons adding additional coding sequence to the primer and then amplifying the gene of interest
using an expression vector that has the six codons adjacent to a multiple cloning site adding additional coding sequence to the primer and then amplifying the gene of interest
In the presence of X-gal, bacterial colonies transformed by pUC19 that contains insert DNA will have a color of _____. Multiple choice question. green white yellow blue
white