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Grade 12 Biology > Biotechnology Part 2 > Flashcards

Biotechnology Part 2 Flashcards

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1
Q

what is DNA fingerprinting?

A

a method that identifies an individual based on the patterns formed from the variations in the genetic code

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2
Q

what is tandemly repetitive DNA?

A

Also known as variable number tandem repeats (VNTRs) they are idetical DNA sequences repeated in series (tandem)

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3
Q

short tandem repeats (STR)

A

a subcategory where repeat length is short - most commonly 4 bases long

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4
Q

Polymorphic

A
  • many forms - many alleles - more than the standard 2 alleles
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5
Q

what is a DNA fingerprinting method?

A
  • PCR - takes fragment of interest at a specific loci on the chromosome
  • PCR amplifies the fargment
  • uses gel electrophoresis to separate PCR products by size and visualize the DNA fragmentsk
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6
Q

why are noncoding regions used?

A

coding regions are too similar between individuals and even species

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7
Q

paternity inclusion

A

means he can’t be excluded from being a possible paternity but no CONFIRMATION

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8
Q

gene cloning

A

making multiple copies of a single gene by using in vivo amplification

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9
Q

what are the 3 steps of gene cloning?

A
  1. forming recombinant DNA
  2. transformation (followed by many cell devisions)
  3. selection
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10
Q

recombinant DNA

A

genes from two different sources (often different species) combined into one molecule

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11
Q

episome

A

genetic elements that can exist either as a plasmid or as part of the bacterial chromosome

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12
Q

describe step 1, forming recombinant DNA

A
  • gee of interest is inserted into a bacterial plasmid
  • restriction enzyme digest plasmid and gene of interest so they have similar ends
  • hybridization of matching sticky ends on gene of interest and plasmid
  • DNA ligase seals gene of interest with plasmid
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13
Q

plasmid

A

small, circular self-replicating pieces of DNA with a small number of genes that incorporate themselve into the bacterial chromosome

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14
Q

why are plasmids advantageous?

A
  • not required for bacterial cells to survive in normal conditions
  • when there are stresses, genes on plasmids can confer advantages
  • increases genetic variation
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15
Q

what is a cloning vector?

A

artificially manipulated plasmid into which the gene of interest is introduced

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16
Q

what does the cloning vector contain?

A
  • ori
  • promoter
  • restriction sites / cloning site
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17
Q

what does the ori do in a vector?

A

allows the plasmid to replicate in the host cell

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18
Q

cloning site

A
  • where the gene of interest will be inserted (ligated)
  • where transcription can occur because contains an upstream promoter
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19
Q

describe step 2 - transformation gene cloning

A
  • transforms recombinant DNA into bacterial cell
  • all bacteria multiplies the gene of interest is also replicated
  • alteration of bacterial DNA by uptake of foreign DNA
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20
Q

naked DNA

A
  • DNA not inside a cell
  • donor cell can be dead or nonexistant
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21
Q

what is a natural method of transformation?

A
  • some bacteria have surface proteins that recognize and transport DNA from closely related species
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22
Q

what is an artificial method of transformation?

A

transformation may occur when bacteria is subjected to treatments that make the cell wall permeable

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23
Q

what are specific methods of artificial transformation? Describe them.

A
  • Chemical: Cold CaCl2 treatment followed by heat shocking
  • Eletroporation: cell are shocked with an electric current to create holes in the bacterial membrane
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24
Q

describe DNA amplification in vivo

A

once the bacteria are transformed, they are transferred to and grown in a liquid medium so the total number of bacterial cells and DNA increases

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25
Q

bacterial product: no vector
cause?

A

no transformation or transform without vector

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26
Q

bacterial product: empty vector
cause?

A

transform with unsuccessful ligation

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27
Q

bacterial product: recombinant
cause?

A

transformation and successful ligation

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28
Q

what is selection in gene cloning?

A

indentifying colonies of bacteria containing the recombinant DNA

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29
Q

what is plating in gene cloning?

A

taking a sample of the bacteria and growing them on plates

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30
Q

plates have an agar medium containing…..

A

antibiotics
X-gal

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31
Q

antibiotic resistance gene (ampR)

A

allows cells to be resistance to ampinilin - an antibiotic

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32
Q

how are bacteria selected?

A

bacteria cells that properly transformed also have the antibiotic resistance so they will survive after being exposed to the virus

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33
Q

what does beta galatosidase (LacZ) do?

A

enzyme produced will change a clear substrate called X-gal into a blue product

34
Q

ligation

A

joining different nucleic acids usually involving ligase

35
Q

genome

A

an organisms’ complete set of DNA

36
Q

human genome project

A

an international collaborative research effort to sequence and map all genes in human beings

37
Q

what sequencing approach did Celear Genomics (private company headed by Craig Venter) use?

A

shotgun

38
Q

Describe the shotgun sequencing approach

A

radomly breaking up DNA sequences into lots of small pieces, sequencing fragmets and reassembling them by looking for regions of overlap

39
Q

What sequence approach did PUbliv research institutions worldwide use?

A

small scale shotgun

40
Q

describe the small scale shotgun sequencing approach

A

same as shotgun but markers are used a regular intervals in the genome to make it easier to reassemble the sequence

41
Q

What id Fredrick Sanger develop?

A

the dideoxy termination sequencing method

42
Q

What is a dideoxyribonucleotide (ddNTP)?

A

a ribonucleotide missing two oxygens

43
Q

where are the two oxygens msising from? their location

A

both at the 2’ and 3’ position position

44
Q

How do dideoxynucleotides work?

A

the 3’OH that is needed to react with the phosphate group on the 5’ end of the next nucleotide to form a phosphodiester bond is no longer there so the chain can’t continue

45
Q

What materials are used in sanger sequencing?

A
  • DNAP
  • Primer (radioactive)
  • template DNA
  • dNTPs
  • ddNTPs
46
Q

describe the difference between modern sequencing and the original Sanger method?

A
  • ## 4 differently coloured fluorescent dyes used to label the 4 different ddNTPs in a single test tube so it can be run on one lane on a gel electrophoresis
47
Q

what are the 5 main experiments that determine DNA as the genetic material?

A
  • Miescher
  • Griffith
  • Hammerling
  • Avery, McCarty, MacLeod
  • Hershey, Chase
48
Q

What did Fridrich Miescher discover?

A

DNA

49
Q

What did Friedrich Miescher do in his experiments?

A
  • collected white blood cells from pus
  • lysed cells and isolated nuclei
  • where he found a substance he called nuclein
50
Q

describe nuclein

A
  • present in enery cell type tested
  • high is phosphorous
51
Q

What was the DNA vs Protein debate?

A
  • is DNA or protein the most important hereditary molecule?
  • money on protein since had obvious function and 20 amino acids to be combined in dif ways
  • whereas DNA only 4 in same pattern bc of Levene
52
Q

what did Griffith study?

A

bacterium that causes pneunomia
- streptococcus pneumoniae

53
Q

Describe a Rough colony/bacteria

A
  • benign
  • lacks a protective capsule
    recognized and destroyed by host’s immune system
54
Q

describe the Smooth bacteria

A
  • virulent
  • has a polysaccharide capsule that prevents detection by hosts’s immune system
  • kills the host
55
Q

What did Griffith define as the transformation observed phenomenon?

A
  • change in a cell’s function by the transfer of an unknown substance
56
Q

What is the current definition of the transformation observed phenomenon?

A

a change in genotype and phenotype due to assimilation of external DNA by a cell

57
Q

What species did hammerling use?

A

single cellular green alga Acetabularia

58
Q

what were the 3 distinct parts of the acetabularia?

A
  • foot containing the nucleus
  • stalk
  • cap
59
Q

What were the two hammerling experiments?

A
  • removed the cap vs the foot from the plant and observed what regrew
  • grafted the stalk of one alga type onto the base of another alga type
60
Q

What did Avery determine?

A

that DNA is a hereditary material

61
Q

What question did Hershey and Chase ask?

A
  • which viral component is resposible for reprogramming, DNA or protein?
62
Q

What materials were involved in the Hershey chase experiments?

A
  • bacteriophage
  • radioactive sulfur to label proteins
  • radioacitive phosphorus to label DNA
63
Q

describe the centrifugation of the Hershey Chase

A

supernatent were free phage and phage parts and the pellet was the bacterial host cell

64
Q

what happened in the Hershey chase experiment when radioactive sulfur was added?

A

the supernatent was radioactive

65
Q

what happened when radiactive phosphorous was added?

A

the pellet was radioactive

66
Q

what did Beadle and Tatum propose?

A

that genes were responsible for producing enzymes

67
Q

why was using bread mold a good choice for beadle and tatum?

A

aka neurospora crassa
- simple organism
- short life cycle
- only requires few biological substances so it could grow on minimal media

68
Q

what did beadle and tatum hypothesize??

A
  • must be able to synthesize all the other vitamins and other essential nutrients on their own
  • have enzymes that convert the simple substances into essential nutrients needed for growth
69
Q

what did Beadle and Tatum do in their experiment?

A
  • mutated the genes in the mold
  • looked for mutations that affected the mold’s ability to make essential nutrients
70
Q

what were the three steps in the experimental procedure for beadle and tatum?

A
  • X rays to make mutants
  • selection to isolate the mutants
  • supplementation to identify the mutants
71
Q

describe the first step in the beadle and tatum experiment - making mutants

A
  • treated mold with x rays to mutate the DNA
72
Q

describe the second step in the beadle and tatum experiment - isolating mutants

A
  • looked for mutations where mold grew in complete enriched media and could no longer grow in in minimal media
73
Q

what is a complete media?

A

contains eberything needed for growth without mold bacing to synthesize it

74
Q

what does a complete media contain?

A
  • salt
  • sugar
  • all vitamins
  • all amino acids
75
Q

describe a minimal media?

A

something that only has what is essential for growth

76
Q

what does a minimal media contain?

A
  • salt
  • sugar
  • biotin
77
Q

describe the third step in the beadle and tatum experiment - identifying mutants

A

grew the mutant in minimal media pluts vitamins OR nucleic acid, if the mutant grow it laced the ability to make that nutrient

78
Q

describe a supplemented media?

A

only has what is essential for growth plus one nutrient

79
Q

what are the contents of a supplemented media?

A
  • salt
  • sugar
  • vitamin
  • biotin
  • 1 nutrient
80
Q

what was the result of the beadel and tatum rationale?

A

malfunctioning enzyme was due to mutations in genes

81
Q

what is the one gene, one enzyme hypothesis?

A

the function of a gene is to dictate the production of a specific enzyme

82
Q

why would one gene, one enzyme be inaccurate?

A
  • not all enzymes are proteins
  • one protein may have more than one subunit each one a separate polypeptide
  • some genes do not produce polypeptide
  • one gene may produce multiple polypeptides due to splicing

Grade 12 Biology (30 decks)

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