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Human Genetics > Lecture 2 > Flashcards

Lecture 2 Flashcards

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Biology 101 flashcards
1
Q

Gene cloning is a biological method of

A

Purification

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2
Q

Where does the gene of interest get inserted into

A

Vector

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3
Q

Give an example of a vector

A

Plasmid

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4
Q

Once placed into a vector where does it go next

A

Introduced into a host cell

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5
Q

When the vector replicates what does this allow

A

Allows large amounts of it to be isolated and used for multiple applications

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6
Q

Once a gene has been cloned what can be determined

A

Nucleotide sequence

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7
Q

How is a clone used to determine where the cloned gene is expressed

A

It is used as a probe

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8
Q

Why would you use a clone as a probe

A

Determine where and when the clones gene is expressed

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9
Q

How is gene function determined

A

Over expression or mutation of the cloned gene

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10
Q

What is the gene product

A

Protein

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11
Q

Gene product can be purified for what

A

Enzymes
Antibodies
Protein structure

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12
Q

What is transgenesis

A

Genes that are artificially expressed in other organisms

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13
Q

Name 6 things you would need in a gene cloning toolkit

A

Source of DNA

Polymerase chain reaction

Restriction endonucleases

DNA Ligase

Vector

Host cells

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14
Q

Source of DNA will include

A

Genomic DNA gDNA

Complementary DNA cDNA

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15
Q

Where does cDNA derive from

A

Reverse transcription of mRNA

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16
Q

What is a clone

A

DNA molecule that is identical to the DNA molecule from where it derived from

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17
Q

Does the mRNA have any introns

A

No

18
Q

When can we only run pcr

A

Nucleotide sequence is known

19
Q

Why is it good to use pcr

A

Produce lots of the desired DNA sequence

20
Q

What is restriction endonucleases for

A

To cut DNA

21
Q

Restriction endonucleases recognise what

A

4bp frequent
6bp intermediate
8bp infrequent

22
Q

What is DNA Ligase for

A

Join together fragments

23
Q

What is a vector

A

Where cloned DNA is inserted

24
Q

What are the two ways of thinking of a gene clone

A

Method trying to identify a gene in the first place

As a way of manipulating the gene once it has been identified

25
Q

What are the 3 approached for gene identification

A

Mapping

Homology

Functional complementation of mutants

26
Q

What is a collection of gDNA fragments

A

gDNA library

27
Q

How to make a gDNA library

A

Digest gDNA with restriction enzymes and Ligase it into a vector cut with the same enzyme

28
Q

What are the 2 approaches for creating a gDNA

A

Complete digest and partial digest

29
Q

What is complete digest

A

Is with one or several enzymes, incase the chosen gene contains a restriction site for the enzymes used

30
Q

What is partial digest

A

Is with frequent cutting enzyme. This creates overlapping of fragments.

31
Q

After the plasmids are inserted into the host what needs to happen next

A

Select the mutants with the plasmids

Screen for hosts that can grow on the medium lacking arginine

Isolate plasmid from the colony

Sequence the DNA fragment

32
Q

When a plasmid has a fragment inserted what is this called

A

Recombinant DNA

33
Q

What do we use the library for

A

Isolate genes

Identify a full sequence

Identify a close related species

34
Q

How can we get a probe

A

It could be a fragment of the target gene

Or it could be a gene from a different species that share the same gene

35
Q

After gene manipulation what do you do next

A

Analyse the cloned gene sequence

Analyse gene expression profile

Analyse gene function

Produce recombinant protein

Make transgenic organisms

36
Q

Why do we analyse the gene expression profile

A

Determine where and when the gene is expressed

37
Q

Why do we analyse gene function

A

Examine phenotype

Over express the gene

Mutation in the genes

Associate it with certain diseases

38
Q

Why do we produce recombinant proteins

A

Raise antibodies for gene products

39
Q

Why do we make transgenic organisms

A

So we can express the gene to give some desired trait

40
Q

What do pcr primers do

A

Used to incorporate restriction enzyme sites into the gene

41
Q

What must be used to produce a recombinant protein

A

Expression vector

42
Q

What does pcr do

A

Amplifies a particular ba sequence for cloning

Human Genetics (4 decks)

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