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