Chapter 9 Flashcards
Protein-based system used to locate specific nucleotide sequence in vivo; some versions cut DNA as part of gene editing
CRISPR-Cas system
Technique used to determine which colonies on an agar plate contain a given nucleotide sequence
Colony blotting
In vitro procedure used to separate DNA fragments by size
DNA gel electrophoresis
Probe-based vitro technique used to study gene expression patterns
DNA microaray
Single stranded piece of DNA, tagged with an identifiable marker, this is used to detect a complementary nucleotide sequence is called…
DNA probe
In vitro process of determining the nucleotide sequence of a given DNA molecule
DNA sequencing
Technique that uses a fluorescent probe to detect a given nucleotide sequence within intact cells on a microscope slide
Fluorescent In Situ Hybridization
Deliberately altering an organisms genetic information using in vitro techniques
Genetic engineering
Procedure in which a DNA fragment is inserted into a vector and then transfered into another cell, where it will br replicated and passed on to the cell’s progeny
Molecular cloning
In vitro technique used to amplify a specific region of a DNA molecule, increasing the number of copies exponentially
Polymerase chain reaction
DNA molecule created by joining DNA fragments from two different sources
Recombinant DNA molecule
Type of enzyme used in vitro to cut a specific nucleotide sequence in DNA
Restriction enzyme
A DNA molecule, often a plasmid,, that functions as a carrier for cloned DNA
Vector
A process taking place in a test tube or culture dish, outside the human body
In vitro
Process or procedures taking place inside the human body
In vivo
DNA fingerprinting relies on this method/technology
PCR
Genetically engineered microorganisms are used to make medically and commercially valuable proteins, to produce a specific DNA sequence, and as a tool for studying gene function and regulation in this process
Molecular cloning
Uses to ID microorganisms, track spread of disease, ID cancer types and genetic disorders
DNA sequencing
_____ are used to study gene expression
DNA microarray
______ are used to detect colonies that contain a specific DNA sequence.
Colony blots
Colony blots, Fluorescent in shiu hybridization, and microarrys are this type of technology
Probe technology
Type of enzyme that recognizes and cleaves a specific sequence of DNA
Restriction enzyme
Restriction enzymes cut DNA (in vitro / vivo)
Vitro
Restriction enzymes make staggered cuts in the recognition sequence. The resulting fragments have overhangs called
Sticky/ cohesive ends
What does “anneal” mean
Form base pairs
How are restriction enzymes named?
EcoRI
E = genus
co = species
RI = strain
Enzyme that synthesizes DNA from RNA.
Reverse transcriptase
DNA obtained when reverse transcriptase uses RNA template to synthesize a complementary strand of DNA
cDNA
_____ is used to separate DNA fragments by size
DNA gel electrophoresis
Put in to correct order. Cloning process
- Use restriction enzyme to cut that DNA into fragments
- Introduce the recombinant molecule into a host
- Join a fragment of the DNA of interest with the vector, creating a recombinant DNA molecule. The DNA fragment is said to have been inserted into the vector, and the fragment itself is referred to as an insert.
- Use that same restriction enzyme to cut that DNA into fragments.
- Isolate DNA from the organism of interest
5.
1.
4.
3.
2.
Vectors in use today have 3 key features
- Multiple coloning sites
- Selectability marker: a gene that encodes for antibiotic resistance, allows for selectivity in growing
- Second genetic marker: encodes for observable characteristics (differention) in the recombinant cell
A single-stranded RNA as a guide to recognize a particular DNA or RNA sequence within a cell is called
gRNA guide RNA
These two components function in crispr systems
Cas nuclease & gRNA
Type of Cas nuclease used to study gene function
dCas Dead Cas
How is dCas different from a Cas
dCas doesn’t cut the double bonds however it just attaches to them.
Highly automated DNA sequence method that generates huge amounts of data quickly
High-throughput sequencing
The target fragment amplified exponentially in a PCR reaction
PCR product
PCR can be used to ID microorganisms from a small amount of DNA
T or F
T
What enzyme makes the copies in PCR
Taq polymerase
Why does PCR heat and cool DNA
Heat break bonds
Cool allow DNA primers to attach to allow Taq polymerase to make copies
4 components of PCR
DNA
Taq polymerase
Primers
Deoxynucleotides- four nucleotides required to make the DNA
Annealing of 2 complementary strands of DNA from different sources to create a hybrid double stranded molecule
Is used in this process
Hybridization
DNA probe
- Colonies on an agar plate
- Colonies are blotted on to a nylon membrane
- Membrane is soaked in a Alkaline solution to lyse and denature DNA
- Probe is added that binds to protein of interest
- By location of where the probe has bound Colonies that have DNA of interest can be ID
Describes which process
Colony blotting
What does FISH bind to (Type of molecule)
rRNA
Fluorescent in situ hybridization is used to observe individual cells that contain…..
A given nucleotide sequence
To obtain Eukaryotic DNA without introns, _____ is used to make cDNA from an mRNA template
Reverse transcriptase
Scientists use genetically modified ______ of CRISPR systems to modify and study gene function
Cas nuclease
______ can be used to track disease spread and trace outbreaks
Whole genome sequencing (WGS)
______ allows for faster, less-expensive sequencing
High-throughput sequencing
_____ uses cDNA to determine the sequence of RNA
RNA-Seq
Contains 10s or 100s of thousands of oligonucleotides that each function in a manner analogous to a probe.
DNA microarrays
Which method is used to generate a DNA library
Molecular cloning
All of the following are found in what
Origin of replication
Gene encoding resistance to an antibiotics
Multiple-coloning sites
LacZ gene
Vector
Means of distinguishing cells that have taken up recombinant molecules
LacZ
The function of dCas molecule along with gRNA is most closely associated with which molecule
Restriction enzyme
DNA probe
Reverse transcriptase
Ligase
Vector
DNA probe
High-throughput DNA sequencing rarely makes mistakes?
T or False
False
PCR generates fragments of distinct size even when an intact chromesome is used as a template. What determines the boundaries of the amplified fragments
Site to which the primers anneal
Genetically Enginered Bacteria
Restriction enzymes cut out DNA at specific _______ and this forms______
Recognition sequences
Restriction Fragments
Restriction fragments transfered into a new cell must _____ with the DNA of the recipient cell
Replicate
Fragment of DNA with the gene inserted into a plasmid will form…
Recombinant molecule
Part of the DNA that is transfed is called…
Insert
Plasmid carrying the insert is called
Vector
Gene fusion
Gene being studied fused to a ____
Reporter gene
How do you know if gene fusion was successful
Reporter gene fluoreses
Genetically engineered eukaryotes Produce (what are they called) _____
Transgenic Organism
Bacteria or yeast can transfer DNA to plants this is called…
GMO
Nucleic acid hybridization
Is the process of what?
Bringing 2 (strands) of DNA together to see if they match
Nucleic acid hybridization 2 steps
Denature & Annealing
In Annealing / Hybridization we use ______ to locate area of DNA where gene is located.
can be radioactively or Fluorescently tagged to make ID easier
Probe
_____ are uses to study gene expression
DNA microarrays
Colony blots are used to detect colonies that contain a specific DNA sequence; fluorescence in situ hybridization is used in in the same manner
Is this type of technology
Probe technology
Restriction enzymes cut DNA strands at ________ this generates ______
Recognition sequence
Restriction fragments
_____ seperate DNA fragments by size
DNA gel electrophoresis
_______ can locate and alter specific sites in DNA
CRISPR systems
Cas9 (DNA-cutting enzyme) used for gene editing
Uses ________ to recognize specific DNA sequence
Single-stranded RNA
Microorganisms are _____ for medical, industrial, research use
Plants and animals are ______ to create GMO
Altered
Engineered
In DNA cloning
- Isolate DNA
- Cut with restriction enzyme
- Join _____ (DNA fragment) with ____ (plasmid) to generate recombinant molecule
- Introduce to host
- High-copy-number vector then makes large number of proteins
Insert / Vector
Plant or animal with a cloned gene is ______
Transgenic
Eukaryotic gene cloning
- Isolate mRNA for a particular protein
- Reverse transcribe to single DNA strand (no introns)
- Synthesize complementary strand to form double-stranded ______
cDNA
Generating a Recombinant DNA molecule.
Vector usually a _____ or _____
Has origin of replication, carries coloned DNA
MUST have restriction site(s) where DNA is cut so that ______ can be joined to it
Plasmid or bacteriophage
DNA insert
_______ a gene encoding resistance to an antibiotic
Selectable Marker
Recombinant DNA molecule
Gene such as lac z that encodes for a phenotype to distinguish recombinant plasmid from intact vector is called…
Second genetic marker
Recombinant DNA
Contains the recognition sequence of several different restrictions enzymes
Multiple cloning sites
PCR method
Double-stranded DNA with targe sequence serves as…..
Template
DNA probe locates complementary nucleotide sequence and then anneals through
Hybridization
_____ is commonly used to ID which clononies in a collection contain sequence of interest: ID which bacteria carry a particular gene
Colony blotting
A hybrid gene formed from 2 previously independiente genes. It can occur as a result of translation, interstitial deletion, or chromosomal inversion.
Gene fusion
This method uses gas chromatography to create a “microbial fingerprint” based on fatty acid esters
F.A.M.E
