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Cell Bio > Exam 4 (Topic 16) > Flashcards

Exam 4 (Topic 16) Flashcards

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Biology 101 flashcards
1
Q

Enzymes that cleave phosphodiester bonds

A

Restriction Endonucleases

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2
Q

Enzyme that is made by bacteria only to protect from foreign DNA

A

Restriction Endonucleases

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3
Q

Enzyme that only cut DNA at specific nucleotide sequences by chance

A

Restriction Endonucleases

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4
Q

Puts cut DNA back together

A

DNA ligase

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5
Q

In an electric field DNA moves from the negative electrode to the positive electrode and separates based on size

A

Agarose gel electrophoresis

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6
Q

DNA can be stained with a fluorescent dye to visualize with UV light. DNA bands can then be cut out of gel and isolated.

A

Agarose Gel Electrophoresis

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7
Q

The isolation and amplification of a specific region of DNA

A

DNA Cloning

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8
Q

DNA fragments cut by Restriction endonucleases and put back together with DNA ligase

A

Recombinant DNA

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9
Q

Specialized circular DNA molecules used to transform bacteria

A

DNA Plasmids (recombinant)

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10
Q

Multiple Cloning Site (MCS)

A

Short segment of DNA that allow biologists to insert a piece or pieces of DNA into the region

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11
Q

Origin of replication

A

Tells bacteria to replicate

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12
Q

Selection Gene

A

Code for drug resistance

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13
Q

Can be amplified and purified in bacteria

A

Recombinant DNA

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14
Q

The introduction of DNA into bacteria

A

Transformation

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15
Q

Amplifying DNA fragment in vitro (without the use of bacteria)

A

Polymerase Chain Reaction (PCR)

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16
Q

Can make billions of copies of a segment of DNA

A

Polymerase Chain Reaction (PCR)

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17
Q

Extremely sensitive, can detect a single template in a sample

A

Polymerase Chain Reaction (PCR)

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18
Q

For PCR you need the following materials:

A

Template DNA, Forward and Reverse primers that are complementary to the template DNA, Nucleotides (dATP, dGTP, dCTP, dTTP) to generate new DNA, DNA polymerase

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19
Q

PCR Uses

A

Test for viral infection in a blood sample
Isolate viral genome (usually in the case of HIV)
Forensic medicine

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20
Q

A DNA copy of RNA

A

cDNA

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21
Q

How do you create cDNA

A

Reverse Transcriptase

22
Q

Amplifies the viral genome and detects the amplified DNA on a gel

A

PCR

23
Q

Regions of DNA on all chromosomes that is highly variable and unique to each individual

A

Short Tandem Repeats(STRs) in DNA Fingerprint

24
Q

In order to study the function of a gene we must know

A

its DNA sequence

25
Q

Dideoxy DNA sequencing requires

A 
Deoxyribonucleotide (dNTP)
Dideoxyribonucleotide Triphosphates (ddNTP)--smalle quantity compared to normale dNTPs
1 Primer to start the PCR reaction
Template DNA to be sequenced
DNA polymerase
26
Q

How often is dideoxyribonucleotides incorporated in new DNA

A

Rarely

27
Q

Are missing the 3’-OH and terminate replication of that strand

A

Dideoxyribonucleotides during Chain Termination

28
Q

Old fashion DNA sequencing

A

Do 4 PCR reaction where each reaction contains a single dideoxyribonucleotide in small amounts and regular deoxyribonucleotides. Generates lots of DNA fragments each varying in size where the the dideoxyribonucleotide was inserted. Run aragrose gel of each PCR reaction

29
Q

DNA sequencing today

A

Each dideoxyribonucleotide is labeled with a fluorophore (4 bases = 4 colors). We can now measure the fragment size and using a computer that generates a color coded DNA sequence

30
Q

The appearance or behavior an individual organism or protein (red eye vs. white eye)

A

Phenotype

31
Q

The gene responsible for a given phenotype

A

Genotype

32
Q

Gene function can be determined by identifying

A

Mutant Phenotypes

33
Q

We can alter the genotype with the use of

A

Recombinant DNA technology

34
Q

We can replace a normal gene with a defective gene

A

Gene Replacement

35
Q

We can remove a gene completely from an organism

A

Gene Knockout

36
Q

We can add a new gene to an organism

A

Gene addition

37
Q

Any organism who has had its genome altered in any way

A

Transgenic Organism

38
Q

Injected C. elegans with dsRNA oligonucleotide that can base pair with mRNA sequence would inhibit protein production.

A

1998: Craig Mello and Andrew Fire

39
Q

Craig Mello and Andrew Fire

A

Discovered RNA interference (RNAi) and miRNAs

40
Q

The human genome codes for 400 miRNAs which regulate what?

A

At least 1/3 of all protein encoding genes

41
Q

Transcribed in the nucleus and exported to the cytoplasm

A

miRNA

42
Q

RNA Interference miRNA Process

A
  1. dsRNA binds to RISC complex
  2. Unwinds dsRNA and makes ssRNA
  3. ssRNA/RISC comples binds to complementary sequence on mRNA
  4. ssRNA/RISC complex degrades mRNA
43
Q

Generated from foreign dsRNA

A

siRNA

44
Q

RNA Interference siRNA Process

A
  1. Cleaved by enzyme Dicer into small dsRNA (21 nucleotides)
  2. dsRNA binds to RISC complex which unwinds RNA and makes ssRNA, binds to complementary sequence on mRNA and degrades mRNA
45
Q

Enzyme that cleaves dsRNA into small sdRNA

A

Dicer

46
Q

RNAi in the lab

A

If you know the gene sequence, design siena complementary to the mRNA that will cause the mRNA to be degraded and the protein will no longer be made

47
Q

hen the mRNA is degraded and the protein will no longer be made

A

Gene knockdown

48
Q

Knocking down gene expression

A

Gene knockdown

49
Q

To investigate protein fuction, RNAi can be used to…

A

decrease the amount of protein made in a cell (knockdown)

50
Q

What is responsible for unwinding double stranded RNA and degrading complementary mRNA?

A

RISC Complex

Cell Bio (30 decks)

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