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4U BIO > 7.1 > Flashcards

7.1 Flashcards

1
Q

Molecular Biology

A

The study of the structure and functions of nucleic acids and proteins.

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2
Q

Recombinant DNA

A

A molecule of DNA composed of genetic material from different sources.

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3
Q

Restriction Enzymes

A

Enzymes that can cleave hydrogen bonds and cut DNA.

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4
Q

Restriction Endonuclease

A

An enzyme that cleaves (cuts) the interior of double-stranded DNA in a sequence-specific manner.

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5
Q

Target Sequence

A

A short sequence of DNA that is recognized by a restriction enzyme.

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6
Q

Restriction Site

A

The point on a DNA sequence where the strand is cut.

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7
Q

Sequence Specificity

A

The cuts made by restriction endonucleases are specific and predictable. The same enzyme will cut a particular strand of DNA the same way each time, producing an identical set of DNA fragments.

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8
Q

Restriction Fragments

A

A small segment of DNA generated by cutting a larger piece of DNA with a restriction enzyme.

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9
Q

Staggered Cuts

A

Most restriction endonucleases produce a staggered cut that leaves a few unpaired nucleotides on a single strand at each end of the restriction fragment. These are called sticky ends or overhangs.

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10
Q

Step One of Making Recombinant DNA

A

A restriction endonuclease is selected that can cut both DNA fragments to be combined.

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11
Q

Step Two of Making Recombinant DNA

A

Each piece of DNA is reacted with the restriction endonuclease enzyme to produce cut DNA fragments.

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12
Q

Step Three of Making Recombinant DNA

A

The two cut DNA fragments are incubated with another enzyme, DNA lipase. This enzyme seals the breaks in the DNA, forming covalent bonds between the 2 different fragments.

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13
Q

Gene Cloning

A

The process of manipulating DNA to produce many identical copies of a gene or another segment of DNA in foreign cells.

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14
Q

Plasmid

A

In recombinant DNA technology, a self-replicating, closed circular piece of DNA that can act as a carrier of a gene to be cloned in bacteria .

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15
Q

Step One in Bacteria Gene Cloning

A

A recombinant DNA molecule is produced. A vector must be found which in this case is a plasmid from bacteria. The plasmid must have an origin of replication, carry a gene making it resistant to a certain type of drug, and it must have one or more restriction endonuclease sites.

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16
Q

Step Two Gene Cloning in Bacteria

A

The reaction mixture for producing the recombinant DNA is introduced into the bacteria. The bacterial cells are treated with particular chemicals that make the cell membranes permeable to the DNA.

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17
Q

Transformation

A

In recombinant DNA technology, a process in which a bacterial host takes up a segment of DNA from the environment under particular experimental conditions.

18
Q

Step Three of Gene Cloning in Bacteria

A

The bacterial cells are applied to a Petri dish containing growth media that has been supplemented with the antibiotic ampicillin and a derivative of galactose, X-gal, which causes bacterial colonies to turn blue when the bacteria is broken down by the enzyme coded for by the lacZ gene.

19
Q

Step 4 of Gene Cloning in Bacteria

A

Bacterial colonies containing cells that have the recombinant DNA are identified using the process of elimination with markers.

20
Q

Step 5 of Gene Cloning in Bacteria

A

Cells from the colonies that contain the recombinant DNA are selected and grown in liquid culture to produce a larger population.

21
Q

Step 6 in Gene Cloning in Bacteria

A

The recombinant DNA molecules are isolated and purified from the bacterial cells.

22
Q

Step 7 of Gene Cloning in Bacteria

A

A variety of analysis techniques are used to confirm that the correct recombinant DNA molecule has been made.

23
Q

DNA Amplification

A

The process of producing large quantities of DNA from a sample.

24
Q

Polymerase Chain Reaction (PCR)

A

An automated method for amplifying specific regions of DNA from extremely small quantities.

25
Q

Step One In PCR

A

DNA strand to be amplified is heated to a high temperature to denature the DNA into single strands (95 degrees)

26
Q

Step Two in PCR

A

DNA sample is cooled in the presence of two nucleotide primers; complementary to each 3’ end of the DNA fragment to be amplified (55 degrees)

27
Q

Step Three in PCR

A

DNA sample is heated to 72 degrees-a DNA polymerase (Taq polymerase from thermophiles) begins synthesizing new DNA

28
Q

Step Four of PCR

A

Steps 1-3 are repeated several times to make multiple copies of the DNA

29
Q

Applications of PCR

A
  • evolutionary studies
  • screening for genetic defects
  • crime scene DNA
30
Q

Gel Electrophoresis

A

Tool used to separate molecules according to their mass and charge; can be used to separate fragments of DNA.

31
Q

In gel electrophoresis, shorter segments of DNA move ______ than large segments which move _______.

A

Faster;Slower

32
Q

DNA Fingerprinting

A

A technology used to identify individuals by analyzing the DNA sequence of certain regions of their genome.

33
Q

Short Tandem Repeat (STR) Profiling

A

A technology used to identify individuals based on repeating short sequences of DNA in the genome hat vary in length between individuals.

34
Q

DNA Sequencing

A

A method for determining the nucleotide sequence, base by base, of a fragment of DNA.

35
Q

Dideoxy Sequencing

A

A method for determining the sequence of a DNA fragment using dideoxynucleotides which cause termination of DNA synthesis during the procedure.

36
Q

Human Genome Project

A

A project that sequenced the human genome and identified all the genes within it.

37
Q

Site-directed Mutagenesis

A

A method of specifically altering the nucleotide sequence of a region of DNA.

38
Q

What is CRISPR?

A
  • Clustered Regularly Inter-spaced Short Palindromic Repeats
  • A mechanism of gene editing that can be used to alter genetic sequences
39
Q

Properties of CRISPR that make it such a huge breakthrough?

A
  • can be programmed to cut DNA at a certain sequence so possibly a sequence that contains a faulty gene
  • can also insert a new DNA sequence to correct another
  • relatively simple to use
40
Q

Benefits of CRISPR

A
  • can be used to possibly alter the mutations that cause genetic disorders
  • can be used to alter embryos (possibly stopping a genetic disorder before the baby is born)
  • fast and cheap
  • can possibly target cancer cells
41
Q

Ethical Issues of CRISPR

A
  • “designer babies”

- who will have access to the technology (only people with money to pay for it?)

4U BIO (11 decks)

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