Chapter 9: Biotechnology and DNA Technology
site-directed mutagenesis
a targeted and specific change in a gene
a culture of identical cells, each derived from a single "parental cell," is referred to as a(n)
clone
E. coli may pick up a recombinant plasmid from a neighboring E. coli cell by
conjugation
real-time PCR differs from traditional PCR in that real-time PCR amplification is monitored by gel electrophoresis
false
yeasts CANNOT express foreign, eukaryotic genes.
false
the following sequential steps are used to make a recombinant cell. Which of these steps occurs LAST?
grow cells containing vector with the gene of interest
Vaccines using recombinant gene technology
hep B virus, herpes virus, malaria
southern blotting is used to
identify particular sequences of DNA
safety issues for using DNA technology
need to avoid accidental release into the environment; genetically modified crops must be safe for consumption and for the environment
An ampicillin-sensitive culture of E. coli is transformed with a plasmid that contains the gene of interest plus an ampicillin-resistant gene. If it is then plated on an ampicillin-containing growth medium, __________.
only the bacteria with the plasmid will grow
which of the following is an example of a cloning vector
plasmid
_________ ________ __________ is a technique used to quickly amplify specific sequences of DNA.
polymerase chain reaction
recombinant DNA can be introduced into a host cell by any of the following methods EXCEPT
polymerase chain reaction
a good cloning vector
should have a gene or genes that allows for selection of transformed host cells
which of the following statements correctly differentiates biotechnology from rDNA technology?
Biotechnology involves any use of microorganisms or cells to make products, regardless of the means used. Recombinant DNA technology involves methods for handling and modifying DNA and inserting it into different types of cells.
DNA fingerprints are actually
DNA fragments
which of the following might specifically be used as part of a reverse-genetics approach to studying a gene?
RNA interference
gene silencing
Small interfering RNAs (siRNAs) bind to mRNA, which is then destroyed by RNA-induced silencing complex (RISC)
which of the following statements correctly differentiates a genomic library from a cDNA library?
a genomic library contains fragments of the entire DNA in an organism's genome. A cDNA library contains the coding sequences of eukaryotic genes (minus the introns).
genome projects
a large-scale, often multilaboratory effort required to sequence a complex genome
which of the following is NOT a step in southern blotting?
addition of heat-stable DNA polymerase
which of the following is a safety issue related to the use of recombinant DNA?
allergic reactions to components in genetically modified foods; reduction in populations of "desirable" insects due to the use of Bt insecticide; spread of bioengineered traits, such as herbicide resistance, into related weed species
in genetic engineering, antibiotic-resistance genes are usually cloned into vectors to
allow selection for bacteria containing the vector
assume you insert a specific gene into a plasmid and use blue-white screening. you plate the transformed E. coli cells on an ampicillin X-gal medium. Cells that produce blue colonies are
ampicillin resistant but do not contain the new gene
the process of making multiple copies of a DNA molecule is referred to as
amplification
Complementary DNA (cDNA) is made from
an mRNA template
When two DNA pieces cut with the same restriction enzyme are combined, sticky ends will __________.
associate by complementary base pairing and hydrogen bonds
shotgun sequencing
sequences small pieces of genomes which are assembled by a computer
if DNA ligase were NOT used in the creation of a recombinant plasmid,
base-pairing would occur but the sugar phosphate backbone would not be connected
If you insert the gene for Bt toxin from Bacillus thuringiensis into a tomato plant, the resulting plants will __________.
be toxic to insects that eat them
If you put the gene for Bt-toxin from Bacillus thuringiensis into a tomato plant, the resulting plants will __________.
be toxic to insects that eat them
the use of microorganisms, cells, or cell components to make products such as hormones, antibiotics, food, or vaccines is known as
biotechnology
which of the following statements correctly differentiates biotechnology and rDNA technology?
biotechnology involves any use of microorganisms or cells to make products, regardless of the means used. rDNA technology involves the specific use of molecular modifications in microorganisms or cells, in which a gene from one cell is inserted into another cell, altering the recipient cell to make some desired product.
for the introduction of a genetically modified plasmid into E. coli,
calcium chloride and heat shock can be used
to express a human gene in a bacterium, cDNA must be made because bacteria
cannot remove introns
vectors
carry new DNA to desired cell; must be able to self-replicate; plasmids and viruses can be used as vectors; shuttle vectors exist in several different species and can move cloned sequences among various organisms
transformation
cells take up DNA from the surrounding environment
recombinant DNA technology is used for all of the following EXCEPT
culturing unknown organisms
restriction enzymes
cut specific sequences of DNA; destroy bacteriophage DNA in bacterial cells; methylated cytosines in bacteria protect their own DNA from digestion; create blunt ends or staggered cuts known as sticky ends
in nature, the function of restriction enzymes is to
destroy bacteriophage DNA
a plasmid that has been cleaved with EcoRI (one) can recombine with another plasmid that has been
digested with EcoRI
disadvantages when making a gene product
e.coli - produces endotoxins and does not secrete its protein products mammalian cells - harder to grow
pros for using E.coli to make a gene product
easily grown and its genomics are known, divides about every 20 mins
electroporation
electrical current forms pores in cell membranes
special considerations must be taken when using bacteria to produce a eukaryotic protein. What is the cause for this additional difficulty?
eukaryotic genes contain introns, which prokaryotic cells cannot remove
for Agrobacterium tumefaciens to be used to introduce foreign DNA into a plant cell, that DNA must first be
inserted into the T-DNA region of the Ti plasmid of A. tumefaciens
RNA interference
inserts DNA encoding siRNA into a plasmid and transferred into a cell
medical uses for dna technology
insulin, human growth hormone, vaccines
which of the following is NOT true of the polymerase chain reaction (PCR)?
large amounts of DNA must be isolated from the source organism
microinjection
like a tiny shot, in vitro fertilization (ivf) occurs like this
subunit vaccines
made from pathogen proteins in genetically modified yeasts
in genetic engineering, antibiotic resistance genes are often cloned into a vector to
make direct selection of a clone possible
mutation
mutagens cause mutations that might result in a microbe with a desirable trait
polymerase chain reaction (PCR)
process of increasing small quantities (amplifying) of DNA for analysis; used for diagnostic tests for genetic diseases and detecting pathogens; reverse-transcription PCR uses mRNA as a template
in the blue-white screening procedure, bacteria that are transformed with recombinant plasmid and cultured in media containing ampicillin and X-gal will
produce white colonies
which of the following is NOT an advantage of obtaining the protein produce called human growth hormone by recombinant DNA technology rather than extraction from cadavers?
production of endotoxins
which of the following is NOT an advantage of obtaining the protein product human growth hormone by recombinant DNA technology rather than extraction from cadavers?
production of endotoxins
protoplast fusion
removing cell walls from two bacteria allows them to fuse
___________, useful in recombination DNA technology, are bacterial enzymes that recognize and cut specific sequences of DNA
restriction enzymes
which of these statements is true for restriction enzymes?
restriction enzymes are useful in genetic engineering when they make staggered cuts in DNA
selection
selecting for a naturally occurring microbe that produces a desired product
tools of biotechnology
selection, mutation, site-directed mutagenesis
the shotgun sequencing technique is used to
sequence entire genomes
the human genome project
sequenced the entire human genome
plamids that can exist in disparate species such as a bacterium and a plant cell are called ________ vectors, and they can be used to transfer cloned DNA from one type of organism to another
shuttle
which of the following can be used as vectors to genetically modify cells?
shuttle vectors, plasmids, and viruses
the following steps are necessary to clone eukaryotic genes in bacteria. what is the third step?
splice exons together
therapeutic applications of DNA technology
subunit vaccines, gene therapy, gene editing, gene silencing, RNA interference
Genetic technology has enabled screening for a variety of genetic conditions, and use of this technology is becoming more widely available. Which of the following is likely to become an important issue that will need to be addressed?
the need for legislation to protect the privacy of individuals' genetic information
sequencing a genome directly provides
the order of nucleotides in a genome
metagenomics is
the study of genetic material directly from environmental samples
biotechnology
the use of microorganisms, cells, or cell components to make a product; food, antibiotics, vitamins, or enzymes - utilisation of the following for the benefit of mankind - organisms, part of organisms, biological processes
during the southern blotting technique, what is the purpose of transferring the DNA fragments from the gel to a nitrocellulose filter?
this step attaches the DNA fragments to a permanent substrate, which then can be probed
which of the following is NOT a purpose of genetic modification?
to remove antibiotic resistant plasmids from bacteria
gene therapy
to replace defective or missing genes
if a recombinant plasmid is put in solution with E. coli, the bacteria may pick up the plasmid by
transformation
the basic steps to genetically modify a cell are listed below. which step would come LAST?
transformation
the introduction of external pieces of "naked" DNA from solution into a cell is referred to as
transformation
ways to insert foreign DNA into cells
transformation, electroporation, protoplast fusion, gene gun, and microinjection
RNA interference (RNAi) can be used to silence a gene, and it holds promise for treating certain genetic disorders and viral infections
true
if a foreign gene inserted into a plasmid inactivates the b-galactosidase gene, a bacterium containing that plasmid would form blue colonies on X-gal medium
true
when bacteria are genetically modified to produce a protein product, technical difficulties arise because that product is NOT always secreted from the cell.
true
all of the following are benefits and improvements made possible by recombinant DNA technology EXCEPT
use of baker's yeast to produce bread
gene editing
using CRISPR to correct genetic mutations at specific locations
the human protozone project
will map proteins expressed in human cells
pros for making a gene product
yeast cell - easily grown and has a larger genome than bacteria, expresses eukaryotic genes easily e.coli - easily grown and its genomics are known, divides about every 20 mins plant cells and whole plants - expresses eukaryotic genes easily, easily grown, large-scale, and low-cost mammalian cells - expresses eukaryotic genes easily, can make products ofr medical use