Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

MOLECULAR BIO > CH.2 PPT > Flashcards

CH.2 PPT Flashcards

You may prefer our related Brainscape-certified flashcards:
Biology 101 flashcards
1
Q

what is a cloning vector

A

DNA (often derived from a plasmid or a bacteriophage genome) that can be used to propagate an incorporated DNA sequence in a host cell.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

what is genetic engineering

A

manipulation of genes using biotechnology

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

what is an endonuclease and what can it be specific to?

A

nuclease that cleaves phosphoester bonds WITHIN a nucleic acid chain
- may be specific for RNA or for single stranded or double stranded DNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

what is an exonuclease and what can it be specific to?

A

nuclease that cleaves phosphoester bonds one at a time from the end (OUTSIDE) of the polynucleotide chain
- may be specific for either the 5’ or 3’ end of DNA or RNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

what is a restriction endonuclease (* slide)

A

an enzyme that recognizes specific short sequences of DNA and cleaves the duplex (at the target site or elsewhere)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

what does a Type 1 restriction endonuclease do (* slide)

A

cut randomly (no specific target site), far from the recognition sequence (> or equal to 1000 bp)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

what does a Type II restriction endonuclease do (* slide)

A

cut at a palindromic (sequence reads same forward as backwards) recognition site, most common.
- has a recognition sequence and a cutting site

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

what does a Type III restriction endonuclease do (* slide)

A

recognize 2 non-palindromic sequences and cut 20-30bp after the recognition site.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

what does a Type IV restriction endonuclease do (* slide)

A

recognize modified (typically methylated) DNA.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

what does a Type V restriction endonuclease do (* slide)

A

use guide RNAs to target specific non-palindromic sequences (CRISPR-Cas9 system).

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

which of the following sequence is inversely palindromic
a. 5’ GCATGC 3’
b. 5’ GCAACG 3’
c. 5’ GCAT 3’
d. 5’ GCAACGC 3’
e. 5’ GGAAAAGG 3’

A

a. 5’ GCATGC 3’

because the next sequence would be
3’ CGTACG 5’ which is the inverse of the sequence above

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

what is cloning (* slide)

A

making an identical copy

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

what does cloning a fragment of DNA require? (* slide)

A

a specially engineered vector (plasmid) and an insert

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

what is the most common vector (* slide)

A

plasmid

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

what is a plasmid (* slide)

A

circular DNA molecules that can be copied in bacteria (following transformation)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

what is recombinant DNA (* slide)

A

DNA molecule that has been created by joining together (ligation) two or more molecules from different sources.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

what is a multiple cloning site (MCS) (* slide)

A

sequence of DNA containing a series of tandem restriction endonuclease sites used in cloning vectors for creating recombinant molecules.

18
Q

what is the diagram for a basic cloning vector look like? DRAW IT OUT TO MATCH THE SLIDE

A

(Amp (green) –> pMB1 ori –> plac | LacR binding site | MCS |lacZ –>

19
Q

T/F: not all plasmids will incorporate the insert following ligation

A

TRUE

20
Q

what is transformation (* slide)

A

similar to transfection, acquisition of new genetic material by incorporation of added exogenous, nonverbal DNA

21
Q

T/F: not all plasmids will be incorporated into the bacteria (* slide)

A

TRUE

22
Q

vectors contain an ampicillin resistance gene. what is this? what does it select? (* slide)

A

only bacteria containing a vector (plasmid) grow in the presence of ampicillin (antibiotic) - created colonies and is the
- selection of transformed bacteria

23
Q

what does the lacZ gene in a vector do and what does it select for (* slide)

A

allows for screening (when bacteria are grown in the presence of X-gal).
- selection for transformed bacteria containing recombinant DNA (plasmid + insert in MCS)

24
Q

plasmid + E.coli makes b-galactosidase, which cleaves X-gal and makes it what color? what does this color indicate? (* slide)

A

BLUE = this means the lacZ gene is intact and there is nothing inserted at MCS

25
Q

plasmids with an insert disrupt the lacZ gene such that a functional b-galactosidase, thus creating a bacterial colony that is what color? what does this color mean (* slide)

A

WHITE = contain both the plasmid and the inserted DNA within the plasmid which is what we want

26
Q

iCLICKER: in a plasmid vector, the purpose of the lacZ gene is

A

an indicator that the plasmid is recombinant, containing a DNA insert at the MCS

27
Q

what are 2 methods that exist to introduce DNA into diff target cells? which of the 2 is the most common (* slide)

A
  1. transfection (most common)
  2. transduction: bacteriophage (virus) will inject DNA into the cell
28
Q

what are reporter genes (* slide)

A

used to measure promoter activity or tissue-specific expression or cellular localization

29
Q

what are promoters (* slide)

A

elements that drive the expression of a gene (gene is NOT expressed w/o a promoter)

30
Q

what are 2 things reporter assays indicate? (* slide)

A
  1. if a gene is expressed in the cell or organism
  2. how expression is affected by stimuli
31
Q

what are expression vectors (* slide) is it constitutive?

A

contain promoters that allow transcription of any cloned gene (gene is inserted AFTER the promoter)

YES. Constitutive = always on/protein constantly expressed

32
Q

what are expression vectors used to understand? (* slide)

A

the function of a gene in any cell type/organism

33
Q

what does inducible mean

A

turned on/off by inducer

34
Q

iClicker: if you wanted to measure the promoter activity of a specific gene of interest, which of the following is NOT an important feature of the vector?

a. Multiple cloning site (MCS)
b. Inducible promoter system
c. antibiotic resistance gene
d. blue/white selection system
e. reporter gene (e.g., luciferase, GFP)

A

b. Inducible promoter system

35
Q

what is transgenics?

A

organisms created by introducing DNA prepared in test tubes into the germline

36
Q

in trans genetics the DNA can be inserted…

A

into the genome or exist in an extrachromosomal structure

37
Q

the Cre/lox system is widely used to make inducible… (* slide)

A

knockouts and knock-ins

38
Q

an endogenous gene can be replaced by…

A

a transfected gene using a homologous recombination

39
Q

what is a knockout (KO) (* slide)

A

A process in which a gene function is eliminated, usually by replacing or removing most of the coding sequence. “Complete erasure or inactivation of the target gene”; applicable at DNA level.

40
Q

what is a knock-in (* slide)

A

A process similar to a knockout, but foreign genetic material is incorporated (more subtle). One-for-one substitution of DNA sequence information in a genetic locus or the insertion of sequence information not found within the locus.

41
Q

what is a knock-down (* slide)

A

A process to suppress the expression of a gene; applicable at the RNA level (more subtle than knockout)

42
Q

iClicker: which of the following is not a good feature of a basic plasmid vector
a. Multiple cloning site (MCS)
b. Telomeres
c. Antibiotic resistance gene
d. origin of replication
e. blue/white selection system (lacZ)

A

b. Telomeres

MOLECULAR BIO (6 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions