Molecular Genetics Flashcards

0
Q

Hershey chase experiment

A

Solidified the concept of DNA being hereditary material

T2 pages grown in medium containing radioactive phosphorus (only found in DNA) and results were that the infected bacteria when centrifuged, had the p32. Sulfur was not

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1
Q

Griffiths experiment

A

Trying to develop a strain for bacteria

Two kinds, R (rough w/ no polysaccharide coat) and S (smooth and with)

After mixing R strain with heat killed S strain, he noticed the mice died

Conclusion: transformation is defined as a change in genotype and phenotype due to assimilation of external DNA

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2
Q

Chargaff’s rule

A

Amount of adenine=amount of thymine

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3
Q

DNA copying direction

A

New nucleotides are added to the 3’ carbon of the sugar

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4
Q

Enzymes in DNA replication

A

Helicase: unwind/separate DNA

Single strand binding proteins: holds strands apart

Primase: starts copying process by adding an RNA primer

DNA Polymerase: adds DNA nucleotides beginning at primer

Polymerase II: removes primers and replaces with DNA

Ligase: joins strands together

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5
Q

Function of DNA

A

DNA codes for genes which are expressed in an organisms genotype and phenotype

Follows three steps:

  • Transcription
  • RNA processing
  • Translation
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6
Q

Redundant code

A

A protein can be coded for by multiple three-sequence DNA strands with only a change in the last letter

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7
Q

DNA transcription

A

Occurs in nucleus

RNA polymerase has two duties:

  • unwind DNA
  • make RNA copy of DNA

To make the copy, polymerase begins imitation and attached to the promoter on the template strand, then begins elongation and adds RNA nucleotides, and finally termination so mRNA can detach

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8
Q

RNA processing

A

Occurs throughout many steps:

  • first, a modified guanosine triphosphate attaches to the 5 prime end to make a 5’ cap, which tells ribosome to attach here to begin translation
  • on the other end, a poly a tail is added to keep mRNA from getting hydrolyzed
  • introns or “intervening sequences” are removed from the coding segment by spliceosomes, which are made of “snurps”
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9
Q

Order of segments on RNA strand

A
5' cap
Leader
Start codon
Coding segment (exons)
Stop codon
Trailer (has termination signal)
Poly a tail
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10
Q

Protein synthesis

A

At initiation, a small ribosomal subunit binds to the mRNA at the start codon, AUG. A tRNA molecule with methionine and the anticodon, AUC, also binds to the mRNA at the same place

A large ribosomal subunit attaches to the mRNA at its “p” site which is in the center. So far, an initiation complex had formed

Next phase is elongation, which begins when tRNA molecules from the cell approach the complex and match up with the correct codons located at “a” sites to the right. Up top, peptide bonds are formed between amino acids in a growing chain that leaves from the “e” site.

Termination begins when any of the three stop codons (UAA, UAG, UGA) enters the p site, after which a release factor binds to the a site causing everything to collapse

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11
Q

Point mutations

A

A change in one or a few base pairs in a gene

Can be divided into Base pair substitutions or base pair insertions/deletions

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12
Q

Base pair substitutions

A

The replacement of one nucleotide and its partner with another pair

Silent mutations: no effect
Missense mutation: a single amino acid is changed in the resulting protein
Nonsense mutation: change creates a stop codon; protein is nonfunctional

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13
Q

Base pair insertions/deletions

A

Also called frameshift mutations because a single addition/deletion can cause a complete shift in the reading frame

Can be disastrous by creating amino acids to make a protein that’s really not useful, or harmless (if it adds three)

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14
Q

Viral structure

A

Protective coat:

  • Capsid (made of protein)
  • Envelope that cloaks capsid and is made from membrane of host cell
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15
Q

Viral genome

A

Double stranded/single stranded are both possible

DNA/RNA are both possible

16
Q

Viral reproduction

A

For example: (lytic cycle)

T4 phage uses tail fibers to stick to receptor sites on E. coli cell

Phage injects DNA through hole

Cells DNA is hydrolyzed

Phage DNA directs production of phage proteins and DNA

To leave the phage directs production of lysozyme which splits open the cell

In the lysogenic cycle, phage integrates into the bacterial chromosome, and remains dormant for generations until prophage separates and undergoes lytic cycle

17
Q

Retroviruses

A

Viruses such as HIV that can make DNA from RNA using reverse transcriptase

18
Q

Prion

A

A protein particle that is the cause of brain diseases

Scrapie agent that leads to disease could be a malformed protein

19
Q

All about bacteria

A

Chromosome: double stranded circular DNA, located in membrane less “nucleoid”

Plasmids are smaller and more numerous

Asexual reproduction is with binary fission and starts at origin of replication

20
Q

Bacterial DNA exchange

A

Transformation (taking up of naked, foreign DNA from surroundings)

Transduction (phage viruses carry genes from one bacterial cell to another)

Conjugation (direct transfer between two cells temporarily joined)

21
Q

The operon

A

A mechanism that represses expression of genes needed for a pathway when the end product is too high

Tryptophan operon: e. Coli synthesizes tryptophan in five steps, each step is controlled by an enzyme that is coded by a different gene, all of which lie on the same DNA strand and are controlled by a promoter. The operator is a segment after the promoter, and proteins attach here to act as repressors so the operon is turned off. For the repressor to exist in active form, tryptophan needs to allosterically activate it

The allolactose operon functions the opposite way, with a regulatory gene that produces proteins in their active form, and the end product must inhibit them so the operator is free and the operon can resume production

22
Q

Transposon

A

A piece of DNA that can move from one location to another in a cells genome

Two types:

Insertion sequences have the DNA for the transposition, as well as inverted repeats on either side

Composite transposons include more genes than transposase

23
Q

DNA packaging

A

Histones are proteins that provide first level of packaging

Nucleosomes are the combo of histones and DNA

24
Q

Restriction enzymes

A

Allow DNA from two different organisms to be stitched together

DNA ligase must be added to bond each strand together

DNA used is called recombinant DNA

25
Q

Gel electrophoresis

A

A process by which restriction enzymes are used to cut up strands of DNA into different sized pieces (due to unique number of restriction sites)

Under the influence of electric field, DNA fragments move through gel and the smaller fragments move faster

26
Q

Tandemly repetitive DNA

A

Short sequence of DNA repeated many times

10-15% of our DNA

Has a structural role for chromosomes

Can lead to huntingtons disease or fragile x syndrome

27
Q

Interspersed repetitive DNA

A

Scattered randomly throughout genome

25-40% of DNA

Repeated units are much longer

28
Q

Eukaryotic genes in prokaryotes

A

This is a problem as there are introns in the DNA of eukaryotes, which the prokaryote is not able to splice with enzymes, so artificial genes called cDNA are made, which lack introns, so they can be introduced in the bacterial cell

29
Q

Polymerase chain reaction

A

Materials:

piece of DNA
Heat resistant DNA polymerase
DNA primers
Four nucleotides (for creating new DNA)

Steps (at varying temps.)

Heat to separate DNA strands (denaturation)
Cool to allow primers to bind (annealing)
Polymerase extends end of primers (extends)