Chapter 11: Genetic Engineering and Biotechnology
Which of the following germs is used to describe a synthetic DNA fragment?
DNA cassette
The enzyme that covalently links both stands of a vector and inserted DNA in molecular cloning is
DNA ligase
Which objective would be best to use a Southern blot rather than a Northern blot?
Determine if a gene is present in a genome
Engineering a metabolic pathway enables a researcher to use different genes from unrelated organisms. (T/F)
False
Genomic libraries enable the discovery of individual gene(s) involved in a articular function of interest with cloning vectors in an expression host, such as E. coli. (T/F)
False
Green fluorescent protein (GFP) is used for detecting translational activity of a fused protein, whereas lacZ reporters are used to detect transcriptional activity of a fused gene. (T/F)
False
High expression levels of a eukaryotic gene in a bacterium such as E. coli cannot be accomplished due to the presence of introns. (T/F)
False
One problem with both BACs and YACs is that genetic regions of these chromosomes cannot be subcloned. (T/F)
False
The lacZ gene is commonly used as a reporter gene, because its substrate lactose is well known and easily meaursed. (T/F)
False
Which of the following sequences would be cleaved by a type II restriction enconuclease?
GAATTCCTTAAG
Which of the following is NOT a common step in creating a genomic library?
Hybridize DNA sequences to form inserts of a target size range.
What type of vector can replicate and be maintained stably in two (or more) unrelated host organisms?
Shuttle
Some proteins overexpressed at high levels resulting in the formation of inclusion bodies can abolish the goal of producing large quantities of active protein. What could be done to minimize this issue?
Simultaneously produce intracellular chaperonins
Although various codons often d=code for he same amino acid, it is important to choose the codon preferred by the expression host itself. (T/F)
True
Artificially synthesizing DNA strands (e.g. oligonucleotide primers) involves the careful attachment of one nucleotide at a time to an immobilized sequence. (T/F)
True
DNA ligase mediates the insertion of foreign DNA into a vector, but it will only be able to do so if the inserts and vector have matching sticky or blunt ends. (T/F)
True
DNA polymerases from E. coli cannot be used to artificially copy gene sequences with a thermocycler. (T/F)
True
Developing vaccines for humans relies heavily on manipulating and engineering vectors. (T/F)
True
Due to well developed molecular tools and careful screening designs, functional genes can be isolated directly by isolation from the environment rather than cultivating the diverse species in a microbial community. (T/F)
True
If vaccinia viruses were non both immunogenic and relatively benign; they would likely not be favored vehicle for vaccinations. (T/F)
True
In principle, a type II restriction endonuclease with an 8-nucleotide recognition sequence should cut 1 in every 4^8 nucleotide positions. (T/F)
True
Modification enzymes typically methylate specific bases within the recognition sequence to prevent digestion of the nucleotide sequence by restriction endonucleases. (T/F)
True
One fundamental technique of genetic engineering includes the ability to cut DNA into random fragments. (T/F)
True
One important advantage of eukarytotic cells as hosts for cloning vectors is that they already possess the complex RNA and posttranslational processing systems required for the production of eukaryotic proteins. (T/F)
True
One method to circumvent issues with introns when expression eukaryotic gene is a bacterium is t simply clone the mature transcript. (T/F)
True
Regardless of the DNA polymerase used in PCR, such as Taq or Pfu, they all have an inherent inability to perfectly copy the template strand, which means the polymerases themselves occasionally make mutations in the sequences they copy. (T/F)
True
Strong promoters used for genetic manipulation are usually regulated by specific molecules. (T/F)
True
The key steps in cloning a foreign gene into a vector, regardless of the application, involve isolating the insert fragment, ligating the insert into a vector, and transforming it into a host. (T/F)
True
While other types exist, Type II restriction endonucleases are by far the most commonly used enzymes for genetic engineering. (T/F)
True
Which statement is TRUE?
YACs are more likely than BAC's to undergo recombination and rearrangement.
The principe behind a nucleic acid probe design is that the probe itself must contain
a key complementary part of the target gene sequence of interest.
Inserting a kanamycin resistance cassette into a catabolic operon to confirm the gene is essential in degradation I=of a particular compound would involve all of the following EXCEPT
a reporter gene
To verify a gene was cloned into a vector successfully, sequencing the vector as well as _______ are commonly performed.
agarose gel electrophoresis
Which of those below is NOT an important consideration when designing a fusion protein construct?
avoid hybridization of the fusion gene in the artificial contruct
After digesting a DNA sequence, a restriction endonuclease can generate
blunt ends, overhangs, or sticky ends (all of the above)
To discover a catabolic gene cluster, cloning large sequences of approximately 40kbp requires the utility of
cosmids or fosmids
Which of the following is NOT an example of synthetic biology?
developing a novel polyvalent vaccine
At which time period(s) during PCR thermocycling is/are hottest in temperature?
during DNA denaturation
What molecular mechanism/feature does site-directed mutagenesis exploit to introduce a mutation at a specific site?
flanking complementary bound nucleotides permit non-complementary base pairing
Which of those listed below is LEAST similar in what is measured and concluded?
fluorescence in situ hybridization
A poorly immunogenic vaccine often suggests the foreign proteins were not properly recognized by the immune system due to a lack of ______ necessary, which can also be engineered to occur with additional molecular manipulations.
glycosylation
One of the more formidable obstacles to mammalian gene cloning is the presence of
introns
If a protein to be overexpressed is toxic to the expression host, it is best to select and expression vector that
is inducible
Cloning vectors can be distinguished from expression vectors by
lacking a promoter site upstream of the insertion site
What makes eukaryotic transcripts easier to isolate than transcriptase in bacteria?
mRNA is polyadenylated in eukaryotes
Expression vectors are designed to ensure that ______ can be effeiciently _______.
mRNA/ transcribed
A shuttle vector is most useful for
making a foreign protein in a mammalian cell
Polyvalent vaccines using vaccinia virus are highly favored by doctors and physicians but are especially challenging for those who develop them, because
multiple foreign proteins simultaneously synthesized often disrupts each other's activity
If a foreign gene is cloned into an expression host, it is important that the host itself
not produce the protein being studied
A polymerase chain reaction (PCR) copies an individual gene segment in vitro with a(n) ______ primer(s).
pair of DNA
The Ti plasmid is best suited for genetically manipulating
plants
Using a host defective in proteases is likely to be necessary when engineering
production of a small protein
Which construct would be MOST useful in studying translational control?
protein fusion
To estimate the total concentration of a beneficial bacterial species in yogurt, _______ would provide the quickest results.
qPCR
Type II restriction endonucleases
recognize nucleotide sequences that are palindromic
The principle underlying how salmon were genetically engineered to grow faster is the
replacement of inducible to constitutive hormone production
A(n) _______gene is a gene that encodes a protein that is easy to detect and assay.
reporter
The genes encoding lucriferase, green fluorescent protein (GFP), and beta-galactosidae are typically used in cloning as
reporter genes
Detecting a specific protein with an antibody is considered a(n) _______ method
screening
Recognizing pathogens that contain multiple unique which enable the human immune system to recognize just one and mount an effective response has opened the door on development of some vaccines only being
subunit vaccines
Which of the following is NOT a characteristic of a type II restriction endonuclease?
unable to cleave methylated DNA