Ch. 9 Microbiology Assignment Questions

rank the fragments as they would appear from the top of the gel towards the bottom of the gel - 10000bp - 9500bp - 5000bp - 500bp - 150bp

- 10000bp - 9500bp - 5000bp - 500bp - 150bp

genetically engineered bacteria have many uses, including: - DNA production - research - gel electrophoresis - protein production

- DNA production - research - protein production

Please rank the steps involved in the dideoxy chain termination DNA sequencing method from the beginning to the end of the process - DNA template, DNA polymerase, primer, deoxynucleotides and labeled dideoxynucleotides are mixed together - the reaction tubes are incubated at an appropriate temperature, facilitating primer annealing and DNA synthesis - the sample is heated, causing DNA denaturation - DNA fragments are separated by gel electrophoresis - a laser is used to detect the fluorescent dyes of the dideoxynucleotides - the DNA sequence is obtained by determining the order of the colored peaks obtained after laser analysis of the gel

- DNA template, DNA polymerase, primer, deoxynucleotides and labeled dideoxynucleotides are mixed together - the reaction tubes are incubated at an appropriate temperature, facilitating primer annealing and DNA synthesis - the sample is heated, causing DNA denaturation - DNA fragments are separated by gel electrophoresis - a laser is used to detect the fluorescent dyes of the dideoxynucleotides - the DNA sequence is obtained by determining the order of the colored peaks obtained after laser analysis of the gel

if the vector of interest has an ampicillin resistance gene as its selective marker and the lac Z gene as its second genetic marker, what should be added to the media to isolate transformed cells - X-gal - penicillin- beta galactosidase - ampicillin

- X-gal - ampicillin

PCR is used in the following applications: - sequencing target DNA - sequencing target proteins - amplifying target DNA - differential disease diagnosis

- amplifying target DNA - differential disease diagnosis

place the steps involved in colony blotting in the correct order - labeled probe is detected using appropriate method - membrane is soaked in solution to lyse cells and denature DNA - bacterial colonies are transferred to a nylon membrane - probe is added; membrane is inclubated to allow hybridization

- bacterial colonies are transferred to a nylon membrane - membrane is soaked in solution to lyse cells and denature DNA - probe is added; membrane is incubated to allow hybridization - labeled probe is detected using appropriate method

put the following steps in order, when analyzing a sample using FISH - add a labeled probe - incubate to allow hybridization - view sample under fluorescence microscope - wash off unbound probe - chemically treat cells to inactivate some enzymes and make it more permeable

- chemically treat cells to inactivate some enzymes and make it more permeable - add a labeled probe - incubate to allow hybridization - wash off unbound probe - view sample under fluorescence microscope

FISH is commonly used to: - detect specific nucleotide sequences - rapidly identify microorganisms directly - study gene expression in organisms

- detect specific nucleotide sequences - rapidly identify microorganisms directly

DNA gels are often immersed in solutions of ___ after electrophoresis to allow us to visualize the DNA - ethidium bromide - ultra violet light - methylene blue

- ethidium bromide - methylene blue

place the following in order to indicate the naming process for restriction enzymes - first letter of bacterial genus - the order discovered in the bacterium - first 2 letters of bacterial species

- first letter of bacterial genus - first 2 letters of bacterial species - the order discovered in the bacterium

legitimate concerns about genetically modified (GM) organisms include - they always cause allergies - they contain "DNA" - genetic transfer to weeds - possible allergens

- genetic transfer to weeds - possible allergens

which of the following are involved in the naming process for restriction enzymes - genus name - order of discovery - palindrome recognized - discovering scientist - year of discovery - species name

- genus name - order of discovery - species name

put the steps used in microarray analysis in the correct order - identify which spots the labeled cDNAs are bound to using a computer screen - isolate mRNA from organism - add cDNAs to microarray and allow for hybridization - wash off unhybridized - synthesize fluorescently labeled cDNA from mRNA

- isolate mRNA from organism - synthesize fluorescently labeled cDNA from mRNA - add cDNAs to microarray and allow for hybridization - wash off unhybridized - identify which spots the labeled cDNAs are bound to using a computer screen

which of the following are limitations to next-gen sequencing - each genome must be sequenced multiple times to ensure accuracy - it is more expensive than older technologies - this sequencing technology is prone to errors - it requires a large amount of computational power - it can only be used to sequence short strands of DNA

- it requires a large amount of computational power - this sequencing technology is prone to errors - each genome must be sequenced multiple times to ensure accuracy

examples of how recombinant DNA techniques have affected society include its impact on - medical diagnoses - agriculture - forensics - computer engineering

- medical diagnoses - agriculture - forensics

if cells from white colonies when plated on media + x-gal, then the transformed cells contained: - pUC vector insert hybrid - pUC vector with a functional lac Z gene - intact pUC vector - pUC vector with a non-functional lac z gene

- pUC vector insert hybrid - pUC vector with a non-functional lac z gene

cloning a segment of DNA into bacteria provides a readily available source of ___ - virus - protein - nucleotides - DNA

- protein - DNA

in addition to separating DNA, gel electrophoresis can also separate ___ according to size - proteins - ethidium bromide - RNA

- proteins - RNA

what is the role of restriction enzymes in the biotechnology lab - synthesizing new proteins - recombinant DNA technology - repairing mutated DNA - protecting against foreign DNA - cutting DNA predictably into fragments

- recombinant DNA technology - cutting DNA predictably into fragments

which of the following is found in a vector used for cloning - restriction enzyme recognition site - DNA polymerase - restriction enzyme gene - selectable marker gene

- restriction enzyme recognition site - selectable marker gene

which of the following is found in a vector used for cloning? - restriction enzyme gene - restriction enzyme recognition site - DNA polymerase - selectable marker gene

- restriction enzyme recognition site - selectable marker gene

which of the following are needed for the dideoxy chain termination method of sequencing - dideoxynucleotides - template DNA - RNA polymerase - trideoxynucleotides - DNA polymerase - deoxynucleotides - mRNA - primer

- template DNA - DNA polymerase - primer - deoxynucleotides - dideoxynucleotides

put the 3 basic steps of PCR in order - DNA synthesis occurs - primers anneal to denature DNA - template DNA is denatured

- template DNA is denatured - primers anneal to denature DNA - DNA synthesis occurs

which of the following statements about PCR primers are true - the primers are complementary to template DNA - the primers are composed of RNA and DNA - primers are designed to anneal to the template DNA at each end of the gene of interest - only one primer is necessary for any PCR reaction

- the primers are complementary to template DNA - primers are designed to anneal to the template DNA at each end of the gene of interest

E.coli strains are generally used in cloning experiments because they are - their antibiotic sensitivities are known - their genetics are well characterized - they are easy to grow - they cannot be transformed

- their genetics are well characterized - they are easy to grow

why did the NIH form the Recombinant Advisory Committee - to provide new jobs to people within the institute - to provide guidelines for researchers performing genetic engineering - to try to ensure that this technology is not used for malicious purposes such as bioterrorism - to make sure that the public shares in profits from genetic engineering experiments

- to provide guidelines for researchers performing genetic engineering - to try to ensure that this technology is not used for malicious purposes such as bioterrorism

genetic engineering has been used in the production of - vaccinations - transgenic animals and plants - pharmaceutical proteins - designer babies - DNA

- vaccinations - transgenic animals and plants - pharmaceutical proteins - DNA

corn, cotton, and potatoes have been engineered to produce a biological insecticide called ___ toxin

Bt

a collection of clones containing the entire genome of an organism is called a ___ library

DNA

___ is the process of producing a genetically identical population of cells or genes in recombinant DNA technology

DNA cloning

a ___ ___ contains anywhere from tens to hundreds of thousands of DNA spots, each of which is a short DNA fragment that functions as a probe

DNA microarray

by cloning a segment of DNA into bacteria, researchers will have a readily available supply of ___ and ___ for study

DNA, protein

for many routine cloning experiments, strains of ___ are commonly used because they are easy to grow and are well characterized

E. coli

the 3 steps of PCR are repeated over and over to generate copies of a) DNA b) RNA c) protein

a) DNA

a widely used technique used for sequencing DNA is a) dideoxy chain termination method b) fluorescent in situ hybridization c) colony blotting

a) dideoxy chain termination method

why does DNA synthesis stop when a dideoxynucleotide is incorporated into a new strand of DNA a) dideoxynucleotides lack the 3' OH group to which DNA polymerase usually adds the next nucleotide b) dideoxynucleotides are nucleotide analogs that DNA fails to recognize c) dideoxynucleotides lack the 5'OH group to which DNA polymerase usually adds the next nucleotide d) dideoxynucleotides are stop codons and signal to DNA polymerase that synthesis is complete

a) dideoxynucleotides lack the 3' OH group to which DNA polymerase usually adds the next nucleotide

what creates the series of different size DNA fragments that are separated by gel electrophoresis in the dideoxy chain termination method? a) dideoxynucleotides terminate the DNA synthesis process at different locations based on the nucleotide sequence b) denaturation results in DNA fragments of varying sizes that are separated out on the gel c) different restriction enzymes will cut the DNA template between different nitrogenous bases d) primers that start the process at different locations on the test DNA strands

a) dideoxynucleotides terminate the DNA synthesis process at different locations based on the nucleotide sequence

a lab technique for separating DNA fragments according to size by using electricity to force the DNA through a gel like matrix is: a) gel electrophoresis b) DNA sequencing c) western blotting

a) gel electrophoresis

the process of altering an organisms genetic information in vitro is called ___ a) genetic engineering b) random mutagenesis

a) genetic engineering

chymosin or rennin, which is used to coagulate milk in cheese production, is now commonly made by a) genetic engineering bacteria b) isolating it from cow stomachs

a) genetic engineering bacteria

this heat stable DNA polymerase is used in the polymerase chain reaction a) taq polymerase b) reverse transcriptase c) ecoR1

a) taq polymerase

this technique is used to determine which colonies on an agar plate contain a DNA sequence of interest a) FISH b) colony blotting c) DNA microarray

b) colony blotting

recombinant DNA must be introduced into a suitable ___ cell for replication a) vector b) host c) pseudo

b) host

the hormone ___ is a genetically engineered pharmaceutical protein that is used in the treatment of diabees a) bovine somatotropin b) insulin c) chymosin

b) insulin

biotechnology now uses ___ to engineer genetically altered organisms with traits that allow it to produce useful products and solve practical problems a) traditional genetic screens b) recombinant DNA techniques

b) recombinant DNA techniques

although diagrams often only show one or a few molecules when describing how DNA molecules used by molecular biologists work, in reality ___ a) these molecules are much larger than they appear b) they are working with millions of these molecules

b) they are working with millions of these molecules

what is the practical value of DNA probes a) they are important in determining the size of DNA or RNA b) they can detect specific nucleotide sequences in unknown samples c) they can cut DNA into specific fragments for gel electrophoresis d) they are necessary to begin replication or protein synthesis

b) they can detect specific nucleotide sequences in unknown samples

a bacterium that receives a plasmid with a human insulin gene cloned into it will produce a) human insulin b) insulin DNA c) bacterial insulin d) cattle and pig insulin

c) bacterial insulin

the enzyme ___ is used to convert mature, eukaryotic mRNA into a strand of complementary DNA a) ligase b) intronase c) reverse transcriptase d) DNA polymerase

c) reverse transcriptase

the nucleotide sequence of each primer must be ___ to one end or the other of the target DNA so that only a specific stretch of DNA will be amplified

complementary

a technique that has allowed scientists to study the expression of genes in a given cell is called DNA a) antisense therapy b) genomic mapping c) fingerprinting d) microarray analysis

d) microarray analysis

if the purpose of the experiment is to produce the protein encoded by the eukaryotic DNA of interest, then a vector that is ___ should be used a) able to carry a large insert b) not able to replicate in bacterial cells c) optimized for use in yeast d) optimized for transcription and translation of insert DNA

d) optimized for transcription and translation of insert DNA

E. coli must be made 'competent' so the cells can uptake recombinant DNA by the process: a) transduction b) conjugation c) competence d) transformation

d) transformation

role: building blocks of DNA

deoxynucleotides

role: template containing target sequence

double stranded DNA

a single PCR cycle ___ the number of copies of DNA

doubles

in the fluorescence in situ hybridization (FISH) technique, probes are applied to intact cells and observed microscopically for the presence and location of ___

genes

___ engineering has been used to produce vaccines, pharmaceutical proteins, transgenic plants and animals, and DNA

genetic

the Human Microbiome Project uses ___ to determine the biological diversity in the normal microbiota of humans

genomics and metagenomics

a probe can be used to "find" a sequence of interest because it will anneal to its complement in a process called ___

hybridization

the Human ___ Project uses genomics and metagenomics to determine biological diversity in the normal microbiota of humans

microbiome

the newest technologies of DNA sequencing are called ___ sequencing

next-generation

allows plasmid to replicate independent of host chromosome

origin of replication

the "___ ___ ___" is a technique that makes numerous copies of a given region of DNA in a metter of hours

polymerase chain reaction

role: short DNA molecules that determine site of amplification in target DNA

primers

when two or more DNA molecules are linked together, the resulting molecule is called ___ DNA

recombinant

a(n) ___ gene, such as green fluorescent protein (GFP), can be fused with a gene of interest so, when expressed, the gene product will be visible

reporter

gene fusions are made by joining a gene of interest to a ___ gene in order to study how that gene is regulated

reporter

allows vector to be cut so that gene of interest can be inserted

restriction enzyme recognition site

allows for elimination of bacteria cells that do not contain recombinant molecule or vector

selectable marker

DNA ___ involves determining the exact order of nucleotides in a fragment of DNA

sequencing

role: enzyme that copies target DNA sequence

taq polymerase

a plant or an animal into which a cloned gene has been introduced is called a(n) organism

transgenic

T/F: DNA microarrays can be used to detect specific nucleotide sequences in an organism whose genome has not yet been sequenced

true

T/F: before PCR can be carried out, the nucleotide sequence at each of the gene of interest must be determined

true

T/F: if a gene is cloned into a high copy number plasmid, the recipient cell can potentially make large amounts of that protein

true

a high copy number ___ can be used to express human genes in bacteria

vector