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Genetics 1 - Moenkhe > chapter 19 > Flashcards

chapter 19 Flashcards

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Biology 101 flashcards
1
Q

what is rna replication

A
  • use of enzyme to create a complementary copy of rna
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2
Q

what is dna replication

A
  • geneoems dna is copied in cells
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3
Q

what is transcription

A

copying dna into rna using rna polymerase( main enzyme)

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4
Q

what is translation

A

genetic information from rna (mRNA) into amino acids
- protein synthesis

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5
Q

what is gene regulation

A

the process of controlling which genes in a cells DNA are expressed

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6
Q

what is molecular genetic techniques

A

techniques used to locate,analyze, alter, sequence study and recombine

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7
Q

what are 4 key innovations

A
  • recombinant dna tech
  • polymerase chain reaction
  • DNA sequencing technology
  • genome edititing methods
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8
Q

Define recombinant DNA technology

A

Set of molecular techniques for locating, isolating,altering and studying dna segments

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9
Q

Define restriction enzyme (restriction endonuclease)

A

Recognize specific nucleotide sequences in dna and make double stranded cuts at those seqauences(restriction sites)

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10
Q

restriction enzyme is produced naturally by…

A

bacteria and are used in defense against viruses

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11
Q

how does a bacterium protect its own DNA from restriction enzyme

A

adding methyl group to the dna

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12
Q

Describe the origin of restriction enzymes and how their names are derived

A

from bacteria and their name is derived from the name of the restriction enzyme being used

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13
Q

how are restrictin enzyme name derived

A
  • begins with abbrev. of the bacterial origin
  • first 3 signify the bacterial enzyme, the 4th letter is the strain
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14
Q

Describe the 2 types of cuts restriction enzymes can make (Fig 19.2a)

A
  • HindIII
    -PvuII
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15
Q

what does restriciton enzyme hindIII do

A

cuts the sugar phosphate backbone of each strand and cuts staggered cuts in dna producing single stranded cohesive ends

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16
Q

what does restriction enzyme pvuII do

A

PvuII cuts both strands of dna straight across producing blunt ends

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17
Q

when are restriction enzymes used

A

Restriction enzymes are used whenever dna fragments must be cut or joined

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18
Q

how can restriciton enzyme create recombinant dna

A

In a restriction reaction dna is placed in a small tube with the buffer solution and a small amount of restriciton enzyme then it is heated and the enzyme will have then cut the appropriate restriction sites

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19
Q

how can gel electrophoresis separate dna fragments by size

A

uses an electric current to separate different sized molecules of DNA in a porous sponge like matrix

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20
Q

what charge is dna

A

negative

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21
Q

where does the dna migrate in electrophoresis

A

toward the positive charged electrode

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22
Q

in gel electrophoresis what moves more quickly through the gel

A
  • shorter strands move more quickly than longer strands so the fragments are arranged in order of size
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23
Q

what is CRISPR-case9

A

technology that allows scientists and medical researchers to edit parts of the genome by adding or altering DNA sequence

24
Q

scientists have engineered crRNA from what

A

bacteria with a sequence that is specific to the target sequence of interest

25
Q

the crRNA sequence pairs with what

A

CAS9 protein

26
Q

what does CAS9 do

A

cuts dna at a specific location in the genome

27
Q

steps for CRISPR-cas9 genome editing

A
  1. crRNA from bacteria that is specific to the sequence of interest
  2. other part of crRNA has a sequence that pairs with CAS9 protein
  3. complex finds and binds to the target DNA sequence
  4. CAS-9 makes double stranded cuts in the target DNA sequence
  5. nonhomo ends join ( repair pathway that repairs chromosome breaks)
  6. donar DNA can be inserted into the cell with ends complementary to the break sequence
28
Q

explain how CRISPR- cas9 can be used in gene therapy

A
  • can be used to correct disease causing mutation
  • silence genes
  • prevent or cure disease
  • study gene function
29
Q

what are the different uses of CRISPR-cas 9

A
  • gene therapy
  • treating disease
  • gene modification
  • change gene expression
  • detect pathogens
30
Q

what is a probe

A

Probe: piece of DNA that is complementary to sequence being searched for– has a label on it, ie radioactive so it can be seen

31
Q

what is PCR stand for

A

polymerase chain reaction

32
Q

what is PCR

A

laboratory techniqye used to amplify dna sequences

33
Q

what does replication by pcr require

A

Replication by PCR requires a dna template from which a new dna strand can be copied and a pair of single stranded primers each with a 3- Oh group

34
Q

what are the components of a PCR reaction

A
  1. dna strand for which a new strand can be copied
  2. pair of single stranded primers with OH group to
  3. solution that targets DNA, DNA polymerase and all 4 dNTPS
  4. magnesium ions and other salts that are necessary for the reaction
35
Q

what is a primer

A

short fragments of dna

36
Q

what are the 5 basic components of PCR

A
  1. dna template
  2. dna polymerase enzyme
  3. primers
  4. nucleotides
  5. reaction buffers
37
Q

explain the steps of PCR cycle

A

Step 1: the dna is heated to 90-100 degrees to separate the dna strands
The dna is cooled to 30-65 so that the short single stranded primers can then anneal to their complimnetyar sequences
Step 3: the solution is heated to 72 degrees so dna plymerase can synthesize the new dna strands
Step 4: the cycle is repeated each tume the cycle is repeated the amount of target dna doubles

38
Q

what is gene cloning

A

produce identical copies of a gene
- alter the DNA sequence

39
Q

what is the process of gene cloning

A

main point: place gene into a host genome(bacteria) to be replicated, then the cloning vector carriers forhein gene into host cell
Dna fragment is inserted into a BACTERIAL cell and thecell is allowed to replicate the dna after the cell divides more of the dna fragement is then passed on to the daughter cell and then the cell is lysed to release their dna and the desired fragment is isolated from the rest of the bacterial dna

40
Q

what is a cloning vector

A

stable replicating dna molecule to which foreign DNA fragment can be attached for introduction into a cell

41
Q

what are the 3 elements of a cloning vector

A

Origin of replication;makes sure that the vector is replicated within the cell
Selectable markers: enable cells containing the vector to be selected or identified
Restriction sites into which a dna fragment can be inserted

42
Q

define plasmid

A

Ciruclar dna molecules that exist naturally in bacteria- are commonly used as vectors for coling dna fragments in bacteria

43
Q

how can a foreign dna fragment be inserted into a plasmid

A

Cut the foreign dna and the plasmid with the same restriction enzyme
-insert the DNA into the plasmid using ligase
- transfer the DNA into bacteria
If the restriction enzyme makes staggered cuts in the dna, complementary sticky ends are produced on the foreign dna and the plasmid

44
Q

what is transformation in a plasmid

A

bacterial cells take up dna from the external environment

45
Q
  1. Explain how to use selectable markers to identify recombinant cells (Fig 19.9)
A
  • genes that have resistance are used as a selectable marker
  • a plasmid that contains a fragment of lacz because it contains unique restriction sites into which if there is foreign DNA then B-galactosidase will be produced if not it wont produce it
  • produce identical copies of a gene
45
Q

in transformation only cells with ampR can grow where

A

on plates

46
Q

if there is LacZ that produce b galactose present what color is it

A

blue

47
Q

what color is it if there is lac z that doesn’t produce B galactose

A

white

48
Q

what does expression of a foreign gene mean

A
49
Q
  1. Describe how next-generation sequencing works, specifically Illumina sequencing (Fig 19.19)
A

Have made sequencing hundres of times faster and is less expensive than sanger sequencing
- sequencing in parallel

50
Q

Illumnia sequencing

A

Most widely used sequencing
dNTPs that have a fluorecent tag are attatched
The dnTPs have a terminator group that once incorporated into the growing dna prevents the incorporation of any additional nucleotides

51
Q
  1. Define DNA fingerprinting
A

uses dna for identification

52
Q

what is microsatellites

A

tandem repreated sequence
number differs among individuals
short tandem repeats

53
Q
  1. Describe how RNAi could be used to turn off a gene
A
  • uses double stranded RNA (dsRNA) to target mRNA for degradation
  • temporary turn off gene and observe phenotyp
54
Q

fire and mellow experiment

A
  • created transgenic worms that expression GFP
  • ds unc22 RNA was injected and ds gfp was infected
55
Q

how can we use RNAi to treat human disease

A
  • choloestral ( targer apoB)
  • cancer: target oncogene, multidrug resistance gene
  • hiv: target viral genes

Genetics 1 - Moenkhe (9 decks)

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