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Genetics > Molecular Genetic Analysis and Biotechnology (Lec 15) > Flashcards

Molecular Genetic Analysis and Biotechnology (Lec 15) Flashcards

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1
Q

Techniques used for locating, altering, and studying DNA segments are considered why type of technology?

A

Recombinant DNA technology (genetic engineering)

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2
Q

The molecular genetics revolution refers to what?

A

biotechnology: the use of these techniques to develop new products

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3
Q

Describe the molecular techniques and their respective order used to isolate, recombine, and amplify genes

A

isolate DNA segment or gene from remaining DNA; cutting and joining DNA fragments - restriction enzymes; viewing DNA fragments; locating DNA fragments with southern blotting and probes

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4
Q

How doe restriction enzymes work?

A

recognize and cut DNA at specific sequences

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5
Q

How does a restriction enzyme protect itself form being digested by its own enzyme in bacteria?

A

by adding methyl groups to the recognition sequence

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6
Q

In restriction enzymes, fragments with short, single-stranded overhanging ends are called what?

A

cohesive ends

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7
Q

In restriction enzymes, even length ends from both single strands are called what?

A

blunt ends

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8
Q

What are the two types of ways to view DNA fragments?

A

gel electrophoresis; autoradiography

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9
Q

True or false?
a probe is DNA or RNA with a base sequence complementary to a sequence in the gene of interest and allow one to locate DNA fragments

A

true

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10
Q

Match the blank with DNA, RNA, or protein
Southern blot = ___
Northern blot = ____
Western blot = ____

A

Southern blot = DNA
Northern blot = RNA
Western blot = protein

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11
Q

How are DNA fragments amplified?

A

pcr

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12
Q

define gene cloning

A

amplifying a specific piece of DNA via a bacteria cell

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13
Q

define cloning vector

A

a replicating DNA molecule attached with a foreign DNA fragment to be introduced into a cell

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14
Q

What are three things a cloning vector needs?

A

origin of replication; selectable markers; single cleavage site

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15
Q

Circular DNA molecules from bacteria are called what?

A

plasmids

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16
Q

Synthetic DNA fragments containing restriction sites are called what?

A

linkers

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17
Q

What are selectable markers used for?

A

to confirm whether the cells have been transformed or not

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18
Q

Expression vectors contain operon sequences that allow what?

A

DNA to be transcribed and translated

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19
Q

What does the Bt gene encode for in plants?

A

an endotoxin that inserts into the insect gut cell membrane, paralyzing the digestive tract and forming a pore. The insect stops eating and starves to death

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20
Q

What is Taq polymerase?

A

stable DNA polymerase at high temperature

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21
Q

What does real-time pcr show?

A

quantitatively determines the amount of DNA amplified as the reaction proceeds

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22
Q

List some of the limitations of pcr

A

requires prior knowledge of at least part of the sequence of the target DNA to allow for the construction of the primers; the capacity of pcr to amplify extremely small amounts of DNA makes contamination a significant problem; taq polymerase does not have the capacity to be proofread; the size of the fragment that can be amplified by taq polymerase usually less than 2000 bp

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23
Q

List some of the uses of pcr

A

detecting presence of viruses in blood samples; identify genetic variation; isolate DNA from ancient sources; amplify small amounts of DNA from crime scenes; introduce new sequences into a fragment of DNA

24
Q

a collection of clones containing all the DNA fragments from one source is called what?

A

DNA library

25
Q

What are cDNA libraries?

A

consist only of those DNA sequences that are transcribed into RNA

26
Q

True or false?
When creating a gene library, some clones contain the entire gene of interest, others include part of the gene, and most contain none of the gene of interest.

A

true

27
Q

How are DNA libraries screened?

A

clones of library are plated and then probed

28
Q

The following describe what type of hybridization:

DNA probes used to determine the chromosomal location and to visualize a gene while it is in a cell; FISH

A

Situ Hybridization

29
Q

Isolating genes on the basis of their position on a genetic map, chromosome walking, and chromosome jumping are all aspects of what type of cloning?

A

positional cloning

30
Q

By making probes complementary to areas of ___ between cloned fragments in a genomic library, we can connect a gene of interest to a previously mapped, linked gene

A

overlap

31
Q

True or false?

cystic fibrosis is the most common fatal genetic disease in the united states today

A

true

32
Q

CF is autosomal dominant or autosomal recessive disease?

A

autosomal recessive

33
Q

What is the incidence rate of CF?

A

1 in 3,300

34
Q

What is CF characterized by?

A

chronic lung infections, insufficient pancreatic enzyme production, and increased salt concentration in sweat

35
Q

The CF transmembrane regulator gene encodes for what?

A

an ion channel necessary to maintain fluid balance

36
Q

Some DNA fragments have different restriction sites due to mutation for the same restriction enzyme; this causes polymorphisms within a population called what?

A

restriction fragment length polymorphisms

37
Q

ddNTP lacks a 3’ OH group, which terminates DNA synthesis. What is ddNTP used in?

A

Sanger’s sequencing method

38
Q

How does sanger sequencing with p32 and gel electrophoresis work?

A

synthesis terminates at different positions on different strands, which generates a set of DNA fragments of various lengths, each ending in a dideoxynucleotide with the same base; the fragments produced in each reaction are separated by gel electrophoresis; the sequence can be read directly from the bands that appear on the autoradiograph of the gel; the sequence obtained is the complement of the original template strand

39
Q

How does sanger sequencing with fluorescent labels and capillary electrophoresis work?

A

each of the four ddNTP’s is tagged with a different fluorescent dye; fragments that end in the same base have the same colored dye; products are denatured and the fragments produced by the four reactions are mixed and loaded into a single well on a gel; fluorescent dye on the DNA is detected by laser beam; each fragment appears as a peak on the computer

40
Q

Pyrosequencing is a next-generation sequencing technique. Describe it.

A

DNA is broken up into fragments; adapters containing primer sequences are attached to each fragment, the DNA is made single stranded; each DNA fragment is attached to a bead and surrounded by a drop of solution containing reagents of PCR; the DNA fragment is amplified by PCR; DNA is made single stranded, each bead is forced into a well and layered with sequencing reagents; DNA synthesis takes place within each well; a solution containing a specific dATP flows on the plate; when a nucleotide is added to the growing chain, PPi is released and produces a light-emitting chemical reaction; an instrument records the light emitted by each well; the amount of light is proportional to the number of nucleotides added

41
Q

The following describe what:
microsatellites: STRs variable number of copies of repeat sequences possessed by many organisms; detected by pcr; fragments represented as peaks on a graph

A

DNA fingerprinting (DNA profiling)

42
Q

In regards to DNA fingerprinting and the fragments represented as peaks on a graph, explain the difference between homozygotes and heterozygotes

A

homozygotes: single tall peak
heterozygotes: two shorter peaks

43
Q

Microsatellites, also known as ___ can use pcr primers flanking them to amplify and electrophoresis to identify size variants within and between individuals; each individual as a unique compliment of microsatellietes = a ___ ___

A

STRs; DNA fingerprint

44
Q

This type of genetics begins with a phenotype to a gene that encodes the phenotype

A

forward genetics

45
Q

This type of genetics begins with a gene of unknown function, first inducing mutations and then checking the effect of the mutation on the phenotype

A

reverse genetics

46
Q

In oligonucleotide-directed mutagenesis, the oligonucleotide serves as what?

A

a primer for

DNA synthesis which produces a molecule with a single mismatched pair

47
Q

Define transgenic animal

A

an organism permanently altered by the addition of a DNA sequence to its geneome

48
Q

A mouse that carries an inserted DNA sequence at specific locations is called what?

A

knock-in mice

49
Q

In a knock-out experiment, the phenotype of the knockout mice reveals what?

A

the function of the gene

50
Q

What do siRNAs do?

A

silence genes through a process called RNA interference

51
Q

True or false?
RNAi can be used for the treatment of human disease. It has shown to reduce the levels of ApoB and blood cholesterol in nonhuman primates

A

true

52
Q

What are some examples of pharmaceutical products that harness the power of molecular genetics?

A

human growth factor, insulin, clotting factors

53
Q

What are some examples of specialized bacteria that harness the power of molecular genetics?

A

vitamin B2, C; various amino acids; thickener; xanthan

54
Q

What are some examples of agriculture products that harness the power of molecular genetics?

A

round up ready corn, soybeans, wheat

55
Q

What are some examples of genetic testing that harness the power of molecular genetics?

A

genomic medicine, BRCA 1/2 for therapy decisions

56
Q

What are some examples of gene therapy that harness the power of molecular genetics?

A

Liver-directed lentirviral gene therapy in a dog model of hemophilia B

57
Q

A geneticist is interested in the immune function of mice and induces random mutations in a number of genes in mice and then determines which of the resulting mutant mice have impaired immune function. This is an example of what?

A

reverse genetics

Genetics (15 decks)

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