Upgrade to remove ads
NYCPM; MBG chapter 17
Terms in this set (45)
______________ are inherited differences in DNA base sequences; they are abundant and associated with many genetic diseases
Testing for DNA sequence ________ are more sensitive than others and allow for early detection and __________ identification.
DNA __________ is used to identify family relationships and criminals
DNA techniques used for treatment and prevention of disease include insulin, Factor _____, vacines, and _____ therapy
Techniques include the use of _________ enzymes, cloning ____, PCR, gel electrophoresis, blotting and _______, and gene _____.
Gene therapy involves isolating normal genes and ________ them into diseased cells.
Restriction ___________ recognize 4-6bp and cleaves the bonds in both strands creating restriction fragments with ______ ends.
They usually recognize _____________; sequences that are the same when read in 5' to 3' direction on each antiparallel strand
They are used to ID variations and to make _______ (aka _________) DNA.
Recombinant/Chimeric DNA is created from the joining of the sticky ends of 2 __________ DNA fragments
They are joined together by the protein ____ _______.
DNA produced by reverse transcriptase (RT) is called ________.
This is made when mRNA is isolated and transcribed by _________ _________.
A unique characteristic of this kind of DNA is that fact that it will have no ________, because it was transcribed from mRNA.
In laboratories we can __________ synthesize DNA.
We do this using automated machine that can make oligonucleotides up to ______ base pairs long.
This is useful in experiments meant to _________ DNA, by isolating and _________ the genes.
A common technique for identifying DNA sequences is the use of ________.
ds DNA is melted using alkali substances or _______. The now ss DNA is exposed to a _______ probe that will hybridize (anneal to) a complementary sequence.
Some labels that may be attached to probes include radioactive ____, which can be detected by __________.
How well a probe must complementarily match the DNA, aka the _________, is manipulated by varying the _______ and the concentration of salt.
____ _____________ is a labratory technique that can separate molecules based on size.
It takes advantage of the fact that DNA is ________ charged and can repulsed by similar charges.
The _______ the molecule is the further it will be from starting wells.
Gel electrophoresis can use two different kinds of gels.
___________ gel separate by one nucleotide.
_________ gels separate larger fragments.
The bands can be visualized by using _________ _______ staining solution, or by exposing the gel to ___ light, or with the addition of labeled probes.
Describe how nitrocellulose can be used to detect specific DNA sequences in bacteria.
1.Press the nitrocellulose paper onto an agarplate with growing bacterial colonies on it.
2. Some of the bacteria from each colony will have stuck to the paper. Treat it with alkali substances to disrupt the cells and denature the DNA.
3. Heat the paper to fix the DNA to it.
4. Expose the paper to a radioactively labeled probe solution that is specific for the DNA sequences you are looking for.
5. Wash away unreacted probe.
6.Perform autoradiography and the exposed areas on the X-ray film indicate the presence of the labeled hybridized DNA.
Blot tests are tests run using ____ ___________.
A _______ blot test is a test in which DNA in nitrocelluloseis hybridized with DNA probes.
A ________ blot test is the same but tests RNA with DNA probes.
Meanwhile a ________ blot test is very different in that it tests proteins using antibody probes.
1.What substances are tested in these kinds of blot tests? What type of probes are used?
A. Southern blot test
B. Northern Blot test
C. Western Blot test
2.How are the results visualized?
A. DNA with DNA probe
B. RNA with a DNA probe
C. Proteins with an Antibody probe
1.What is a ddNTP? What is different about them?
Once a ddNTP is incorporated into a necleotide chain, the chain cannot be futher _________.
2.What kind of nucleotides can be used by DNA polymerase during replication/transcription?
1. A dideoxynucleotide-> they lack a 3' hydroxyl group
2. dGTP, dCTP, dATP, dUTP
(stand for deoxy -base name- triphosphate)
DNA sequencing accomplished by replicating the same DNA in ___ separate reactions.
In each reaction you will provide DNA _______, all of dNTPs, and one type of _____.
In DNA sequencing the addition of the ______ will interupt and ends the replication when it is added to the chain.
This creates many different sized DNA ______ that end in the ddNTP.
Using gel electrophoresis with wells for the content of the four separate reactions (one with _____, one with ______, one with _____, and one with _____)
You can analyze how many nucleotides were in each fragment based on the ______ position on the gel, and conclude that the fragment ______ is equivalent to the postion in the normal sequence.
Therefore, if the fragment is the Adenine ____, that position in the DNA is usually an Adenine.
One way to amplify DNA is by _______ DNA.
This is accomplished by attaching foreign DNA to a vector and introducing it to a host.
When the host is a bacterium this is called ___________.
When the host is a eukaryotic cell this is called __________.
In nature bacteria can be transformed by _________, plasmids, and ________ (plasmids with lambda phage)
Transfection of eukaryotic cells in nature happens when the cells interact with ________, Adenovirus, ____ DNA, or _______.
Retrovirus (example is HIV)
In DNA Cloning both the plasmid and foreign DNA are cleaved with the same ________ ________.
The fragments with complementary sticky ends are _______ together.
A bacterial cell is ________ by the vector and grown into a large culture.
From DNA cloning you can obtain either _______ or _______.
To get DNA you isolate the ________, and cleave them with the orginal restriction enzyme to isolate the cloned DNA.
To get protein you grow the bacterium in conditions that allow the _______ of the gene, and then isolate the protein.
A DNA _______ is a specific collection of DNA fragments.
A _______ _______ is a set of host cells that collectively contain all of the DNA sequence from the genome of another organism
There is two types of these; a _______ version and an abridged _______ version
To clone a specific gene you must develop a _________
As well as screen a cDNA or DNA library to find a ______ cell/organism whose DNA contains that sequence.
__________ _______ ______ (___) allows for the rapid production of very large amounts of specific segments of DNA.
Well suited for amplification of DNA for clinical and forensic test because you only need a ______ amount of the _____ ______.
Polymerase Chain Reaction (PCR)
The first step of PCR is to ______ the DNA in order to separate the strands.
As you cool it you will add ________ that match thebegining portion of the desired portion of DNA.
In PCR, after adding primers you add _______ _______ DNA polymerase.
Then you repeat the heating and cooling procedure for up to ____ cycles.
This will give give you a ___ ________ fold amplification of the original DNA sample.
1 million (10^6)
____________ are genetic variations among individuals
Human DNA is estimated to contain _________ polymorphisms; involving _______ mutations in both coding and non-coding DNA
Certain polymorphisms near or of certain genes are closely linked with, and the suspected cause of, ________ diseases.
Because of this they are often used as a _______ for the diagnosis of diseases
____________ _________ ________ __________ (RFLP) can be detected by using restriction enzymes.
This happens when a point mutation occurs at _________ site of the restriction enzyme.
This mutation will cause the restriction enzyme to either make no ____, or to cut at a unusual _____.
This changes the fragment _______ from the usual. This indicates the presence of a ______. These are often associated with disease.
Restriction Fragment Length Polymorphism
An ________-_________ Oligonucleotide _______ is a 15-20 bp probe that is synthesized to contain a mutation
That region of the genome is amplified by ______.
The amplified DNA and radioactive probe is tested with a nitrocellulose _____ _______ gel electrophoresis.
If the probe latched onto the DNA the person has the ___________. If no probe is found in the genome band, the person is normal.
Mutations can be tested for using PCR by using _______ containg the mutation.
The DNA will only be amplified if the mutation is _______.
Some polymorphisms are based on how many copies of repetitive DNA the patient has in ______ _______ _______ of their DNA.
Each person can have a variable number of these ______ ________ (VNTR).
highly variable regions
To indentify the number VNTR a person has restrcition enzymes that can recognize _________ sites are used and the size of the fragments produced will vary depending on the number of repeats.
The restriction fragment patterns produced on the gels can also be used to identify individual people as accurately as fingerprints-> "_____ ___________"
Individuals who are closely related have __________ patterns.
DNA chips are small chips that are dotted with _________ of pieces of ______-stranded DNA.
Each piece representing a _____ or a gene segment.
One of these chips are incubated with a sample of the patient's ______.
____________ analysis is used to indentify the alleles present in the patient
If a DNA chip is prepared with cDNA (DNA made from copies of the patient's _______) gene _________ can be analyzed.
Gene chips can be used for the diagnsis of __________ diseases as well, based on chip being dotted with __________ microbial oligonucleotides.
Before DNA technology vaccines were made with ______ or _________ infectious agents
Possible dangers included the potential for __________
Now __________ ________ is used to produce antigenic proteins-> this elminates the risk of infecting the patient
First succesful vaccine made this way was for __________ __.
contamination (if patient was immuno compromised or the pathogen was still viable, then you just infected that person)
Labs produce ________ and _________ hormone from human DNA that is inserted into plasmids and used to transform _________.
There is no immune reaction to this product in patient's since insulin is not __________.
HGH used to be isolated from cadaver _________ tissue. So this is much cheaper and easier to produce now.
Insulin is made by tranforming some bacterium with a plasmid containing coding for the __ _____, and some with plasmid containing the ___ ______ of insulin.
Once these products are isolated they are _______ and their ________ are oxidized.
2 ________ bonds are made between the 2 chains.
__________ _________ proteins are produced in mammalian cell cultures.
_________ ______ (involved in clotting) is defective in people with hemophilla. Now available for patients via DNA technology
_______ ________ ________ (TPA) is a protease that converts plasminogen to plasmin.
________ cleaves fibrin, a major component of blood clots
Recombinant TPA is usually administered to patients after a ______ _______ in case it was caused clot that is still in the circulation system.
Tissue Plasminogen activator
___________ can be made for patients with certain types of anemia
Interleukins are used after _______ ________ transplants.
___-__________ can reduce severity of MS episodes.
All of these proteins are made in __________ _______ cultures.
What does SiRNA and miRNA stand for?
What does SiRNA do?
SiRNA-> (Synthetic) Small interfering RNA
miRNA-> (Natural) Interfering RNA
reduces expression of certain genes
microRNA (miRNA) is synthesized in the nucleus as ___-_______.
This is processed by ______ ____, making it a double stranded miRNA.
miRNA incorporates itself with RISC (________ ________ ________ ________) and then can degrade or inhibit specific mRNAs.
RNA-induced silencing complex
_______ _________ is the ultimate cure for genetic diseases.
The goal is to normal _____ into individuals that have abnormal/defective ones.
In gene therapy, __________ were the first vectors they've tried to use. Inserted gene size limit is __kb)
They removed ____, _____, and ____.
_________ can be used as vectors (36kb)-> but their genome won't integrate with a host's so the patient must be exposed __________.
_________ are now being tried as a possible vector.
THIS SET IS OFTEN IN FOLDERS WITH...
NYCPM MBG chp 12
NYCPM MBG Chapter 13
NYCPM MBG chp 14
NYCPM; MBG chp 15
YOU MIGHT ALSO LIKE...
Microbiology A Systems Approach: Chapter 10
Microbiology A Systems Approach: Chapter 10
Mastering Biology Chapter 12
OTHER SETS BY THIS CREATOR
NYCPM; Physiology exam 2 cases
NYCPM; MBG exam 1 Gout lecture essentials
NYCPM; MBG exam 1 FH4 and Clinical supplement
NYCPM: Immunology hour 1
OTHER QUIZLET SETS
Cold War Test Review Cluster Four: Berlin and Cuba
Combo with "AG: Ch 6, 7, & 8" and 2 others
Chapter 10: Armament/Ordinance
unit 7 the small intestine