4.5 DNA Cloning Flashcards

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1
Q

What has our knowledge of DNA biology led to?

A

an ability to manipulate the genes of organisms

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2
Q

What is a genome?

A

the complete genetic makeup of an organism

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3
Q

What is genetic engineering?

A

practice of cloning genes and then using them to alter the genome of viruses and cells

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4
Q

What are the uses of genetic engineering?

A

ranges from producing a product to treating genetic disorders

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5
Q

What is cloning?

A

the production of identical copies of an organism through asexual means

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6
Q

Why are human identical twins also considered clones?

A

because the first two cells of the embryo separated and each became a complete individual

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7
Q

What is gene cloning?

A

the production of many identical copies of a single gene

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8
Q

Why do biologists clone genes?

A
  • want to produce large quantities of the gene’s protein product such as human insulin
  • learn how a cloned gene codes for a particular protein
  • use the genes to alter the phenotypes of other organisms in a beneficial way
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9
Q

What is gene therapy?

A

when cloned genes are used to modify a human

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10
Q

What are transgenic organisms?

A

organisms with foreign DNA or genes inserted in them

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11
Q

What are transgenic organisms frequently used for?

A

to produce a product desired by humans

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12
Q

What do most processes rely on to produce cloned DNA?

A

recombinant DNA technology and the polymerase chain reaction

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13
Q

What is recombinant DNA (rDNA)?

A

contains DNA from 2 or more different sources such as the human cell and the bacterial cell

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14
Q

What do researchers need to make rDNA?

A

vector

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15
Q

What is a vector?

A

a piece of DNA that can be manipulated such that foreign DNA can be added to it

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16
Q

What is plasmid?

A

a common vector that are small accessory rings of DNA from bacteria that are not part of the bacterial chromosome and are capable of self-replicating

17
Q

How were plasmids discovered?

A

by investigators studying the bacterium Escherichia coli

18
Q

What are the two types of enzymes that are used to introduce a foreign DNA into vector DNA?

A

restriction enzyme: cleaves the vector DNA

DNA ligase: seals DNA fragments

19
Q

Why are both DNAs cleaved with the restriction enzyme?

A

so they can have complementary ends that can anneal

20
Q

Where do hundreds of restriction enzymes occur naturally?

A

in bacteria. where they act as a primitive immune system by cutting up any viral DNA that enters the cell

21
Q

Why are they called restriction enzymes?

A

because they restrict the growth of viruses

22
Q

What else can restriction enzymes be used for?

A

as molecular scissors to cut double-stranded DNA at a specific site

23
Q

Why is both foreign DNA and vector DNA cleaved with the same restriction enzyme?

A

a gap exists into which a piece of foreign DNA can be placed if it ends in bases complementary to those exposed by the restriction enzyme

24
Q

What are “sticky ends”?

A

the single-stranded but complementary ends of the two DNA molecules

25
Q

Why are they called “sticky ends”?

A

they can bind a piece of foreign DNA by complementary base pairing

26
Q

What do “sticky ends” facilitate?

A

the insertion of foreign DNA into vector DNA

27
Q

What is DNA ligase used for?

A

to seal the foreign piece of DNA into the vector

28
Q

What do bacterial cells do?

A

take up recombinant plasmids, especially if the cells are treated to make them more permeable

29
Q

What happens when the plasmid replicates?

A

so does the foreign DNA and thus the gene is cloned

30
Q

What are the steps of gene cloning? (4)

A
  1. restriction enzyme cleaves DNA
  2. DNA ligase seals human gene and plasmid
  3. host cell takes up recombined plasmid
  4. gene cloning occurs
31
Q

Describe cloning a human gene.

A
  • human DNA and bacterial plasmid DNA are cleaved by a specific type of restriction enzyme (ie. human DNA containing the insulin gene is spliced into a plasmid by DNA ligase)
  • gene cloning is achieved after a bacterium takes up a plasmid
  • if the gene functions normally as expected, the product (ie. insulin) may also be retrieved
32
Q

What is DNA fingerprinting?

A

a technique that involves treating the entire genome with restriction enzymes

33
Q

Why does each person have a unique collection of DNA fragment sizes?

A

because each person has their own restriction enzyme sites

34
Q

What is gel electrophoresis?

A

where an electrical current is used to force DNA through a porous gel material

35
Q

What is a DNA fingerprint?

A

each person’s unique collection of DNA fragments are separated according to their size during gel electrophoresis

smaller fragments move farther through the gel than larger fragments and results in a pattern of distinctive bands called a DNA fingerprint

36
Q

What is short tandem repeat profiling?

A

the method of choice

STRs are the same short sequence of DNA bases that recur several times as in GATAGATAGATA

37
Q

Why is STR profiling advantageous?

A

does not require the use of restriction enzymes and instead, PCR is used to amplify target sequences of DNA which are fluorescently labelled