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Microbiology > Recombinant DNA Technology > Flashcards

Recombinant DNA Technology Flashcards

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Biology 101 flashcards
1
Q

Intentionally modifying genomes of organisms for practical purpose

A

Recombinant DNA technology

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2
Q

3 Goals of recombinant DNA technology

A

Eliminate
Combine
Create

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3
Q

3 Goals of recombinant DNA technology

-Eliminate

A

Undesirable Phenotypic traits

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4
Q

3 Goals of recombinant DNA technology

-Combine

A

Beneficial traits of two or more organisms

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5
Q

3 Goals of recombinant DNA technology

-Create

A

Organisms that synthesize products human need

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6
Q

Overview of recombinant DNA technology

6

A
  1. Isolate plasmid
  2. Cleave DNA to obtain desired genomes
  3. Isolate gene
  4. Insert gene into plasmid
  5. Insert plasmid (w/ desired gene bacteria)
  6. Culture bacteria for various use
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7
Q

Physical and chemical agents that produce mutations

A

Mutagens

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8
Q

Scientists utilize mutagens to

A
  • Create changes in microbes’ genomes to change phenotypes

- Select for and culture cells with beneficial characteristics

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9
Q

____________ alone can be isolated.

A

Mutated genes

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10
Q

Explain reverse transcriptase

A
  • Isolated from retroviruses (Genome is RNA)
  • Uses mRNA template to transcribe molecule of cDNA
  • Easier to isolate mRNA molecule for desired protein first
  • mRNA of eukaryotes has introns (non-coding regions) removed……. (leaving only coding DNA)
  • Allows cloning in prokaryotic cells
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11
Q

Molecules of DNA and RNA produced in cell-free solutions

A

Synthetic Nucleic Acids

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12
Q

Uses of synthetic nucleic acids

A

Elucidating the genetic code

Creating genes for specific proteins

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13
Q

Identifying codons for a.a

A

Elucidating the genetic code

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14
Q

Synthesize a gene for practical use

A

Creating genes for specific proteins

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15
Q

Synthesizing DNA and RNA “prodes” to locate specific sequences of nucleotides and Synthesizing antisense nucleic acid molecules use

A

synthetic nucleic acids con’t

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16
Q

Prodes are small sequences of nucleic acid which are labeled with fluorescent marker (enables the identification and presence of specific DNA sequences in certain genes)

A

Synthesizing DNA and RNA “prodes” to locate specific sequences of nucleotides

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17
Q

Nucleic acid sequence which binds and interferes with genes and/or mRNA to prevent expressions (research conducted to control diseases)

A

Synthesizing antisense nucleic acid molecules

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18
Q

Bacterial enzymes that cut DNA molecules only at restriction sites (nucleotide palindromes)

A

Restriction enzymes

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19
Q

Categorized into 2 groups based on type of cut

A

Sticky ends

Blunt ends

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20
Q

R.E. stands for

A

Restriction Enzymes

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21
Q

R.E. are named according to

A
  • Genus species
  • Order found
  • Strain
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22
Q

E.Coli strain R has restriction enzyme

A

ECoR1

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23
Q

The same____can be used in different organisms

A

R.E.

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24
Q

After RE cut DNA, ligase can be added to

A

Ligate the DNA strand

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25
Q

Nucleic acid molecules that deliver a gene into a cell

A

vectors

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26
Q

What are some useful properties of vectors?

A

They are small enough to manipulate in a lab
survive inside cells
contain recognizable genetic marker
ensure genetic expression of gene

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27
Q

Vectors include

A

Viral genosomes transposens and plasmid

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28
Q

Growth on antibodic medium indicates that gene

A

was delivered succuesfully

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29
Q

A collectin of bacterial or phage clones

A

Gene libraries

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30
Q

Each clone in library often contains

A

one gene of an organisms genome

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31
Q

Library may contain all genes of a

A

single chromosomes

32
Q

librarys may also contain set of

A

cDNA complementary to mRNA

33
Q

Production of a gene library

A

1- isolate genome (all DNA)
2- cleave genome into fragments using RE
3- insert each fragment into a vector (plasmid)
4- insert each vector into different cells
5- culture cells, then place into tubes (label, store)

34
Q

Copying (amplifying) large number of identical molecule of DNA molecules

A

The polymerase chain reaction (PCR)

35
Q

Critical to amplify DNA in variety of situations

A

Epidemiologists use to amplify genome of unknown pathogen

-amplified DNA from Bacillus anthracis spores in 2001 to identify source of spores

36
Q

DNA specific, dNTPs polymerase enzyme, thermocycler

A

PCR requires

37
Q

Heat DNA `94 C breaks H-bonds between dsDNA, resulting in 2 separate complementary strands

A

Denaturation

38
Q

Mixture is cooled to ~65 C, so primers in a mixture can anneal to a complementary sequence in template strands of DNA

A

Priming

39
Q

Temperature is slightly elevated to 72 C and polymerase enzyme synthesizes each template strand of DNA using dNTPs

A

Extension

40
Q

PCR can be automated using a _______: machine which heats and col PCR mixture. repeats the 3 steps

A

Thermocycler

41
Q

PRC stand for

A

Polymerase chain reaction

42
Q

PCR

A

Hydrogen bonds breaks
Primers anneal to specific DNA sequences
Polymerization occurs, new strands synthesize from ssDNA

43
Q

PCR DNA is replicated exponentially after

A

Sever cycles

44
Q

Gel electrophoresis

A

Separating DNA Molecules

45
Q

Explain Gel electrophoresis

A

Separates molecules based on electrical charge, size, and shape

  • Allows scentists to isolate DNA of interest
  • Negatively charged DNA drawn toward positive electrode
  • Agarose makes up gel; acts as molecular sieve
  • Smaller fragments migrate caster then larger ones
  • Determines soze by comparing migrated to strandards
46
Q

Migration of DNA

A

smallest DNA fragments travel the fastest

47
Q

DNA transferred from gel to nitrocellulose membrane

A

Southern blot

48
Q

Southern blot probes uses to localize ________

A

DNA sequence of Interest

49
Q

what uses southern blots

A

Genetic “fingerprinting”

Diagnosis of infectious disease

50
Q

Used to detect RNA

A

Northen Blot

51
Q

Explain the southen blot technique (4 Steps)

A

Cut DNA using RE and place cut DNA mixture into separate wells

1) use gel electrophoresis to separate fragments
2) Transfer (blot) DNA fragmens from gel to nitrocellulose membrane
3) Add radioactive probes to membrane
4) incubate and observe for presence of specific DNA sequence

52
Q

Goal of DNA technology is insertion of DNA into ____

A

Cell

53
Q

Uptake of exogenous DNA

A

Transformation

54
Q

Viral transfer of DNA from one microbe —>another

A

Transduction

55
Q

Transfer to plasmid to another bacteria

A

Conjugation

56
Q

Artifical methods are

A

Electroporation
Protoplast fusion
Injection

57
Q

Gene gun and microinjection

A

Injection

58
Q

Electrical current —-> holes so DNA enters

A

Electroporation

59
Q

Expose microbes to polyethylene glycol to promote fusion of 2 microbes (Recombinant DNA)

A

Protoplast fusion

60
Q

Beads coats w/ DNA and shot into target tissue

A

Injection - gene gun and microinjection

61
Q

Locating genes and specific nucleotide sequences on a nucleic acid molecule

A

Genetic mapping

62
Q

DNA microarrays used to screen individuals for inherited disease caused by mutatuins

A

Genetic screening

63
Q

Genetic screening can also identify _____________.

A

Pathogens DNA in blood or tissue

64
Q

Identifying individuals or organisms by their unique DNA sequence

A

DNA fignerprinting (pharmaceutical and therapeutic applications)

65
Q

Pharmaceutical and therapeutic application

A

Amplify DNA (PCR)

  • Cut DNA (using RE)
  • -Each individual may have diff. fragment size
  • Run DNA on gen electrophoresis
66
Q

Missing or defective gens replace with notal copies

A

Gene therapy

67
Q

With gene therapy some patients immune system reacts

A

Nehatively

68
Q

Patient specimens can be examined for presence of gene sequences unique to certain pathogens

A

Medical diagnosis

69
Q

Animals cells, tissue, or organs introduced into human body

A

Xenotransplants

70
Q

(Agricultrual applications)

Recombinant plants and animals altered by addition of genes from other organisms

A

Production of transgenic organisms

71
Q

(agricultrual applications)

Gen from salmonella conveys resistance to gyphosate (round up)

A

Herbicide resistance

72
Q

Farmers can kill weeds without killing crops

Agricultrual applications

A

Herbicide resistance

73
Q

(Agricultrual applications)
Scientists have removed gene for salt tolerance and inserted into tomato and canola plants
Transgenic plants survive produce fruit and remove salt from soil

A

salt tolerance

74
Q

(Agricultrual applications)

crops sprayed with genetically modified bacteria can tolerate mild freezes

A

Freeze resistance

75
Q

(Agricultrual applications)
(bt toxin) naturally occurring toxin only harmful to insects
-organic farmers used to reduce insect damage to crops
-gene for Bt toxin inserted into various crop plants

A

Pest resistance

76
Q

(Agricultrual applications)

tomatoes allowed to ripen on vine and shelf life increase (gene for enzyme that breaks down pectin suppressed)

A

Improvements in nutritional value and yield

77
Q

allows cattle to gain weight more reapidly, have meat with lowe fat contect and produce 10% more milk

A

BGH

Microbiology (26 decks)

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