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Life Sciences > Test - 1 > Flashcards

Test - 1 Flashcards

1
Q

What is used to amplify DNA fragment?

A

PCR

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2
Q

What is used to amplify DNA?

A

Thermostable Taq DNA polymerase enzyme

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3
Q

What are the PCR components?

A

DNA sample, Primers, Nucleotides, Taq polymerase, Mix Buffer, PCR Tube

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4
Q

What are the steps of the PCR process?

A

Denaturing(95C), Annealing(55C) and Extension(72C)

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5
Q

Process to seperate DNA

A

Agarose Gel Electrophoeresis is used to seperate DNA based on molecular weight

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6
Q

What is the 3D matrix used in DNA seperation?

A

Agarose

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7
Q

what electrode does DNA move to during electrophoresis?

A

Negative to positive

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8
Q

What is the stain used during separation of DNA and what light is used to observe it?

A

Ethidium Bromide and Ultraviolet Light

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9
Q

Method to separate proteins based on molecular weight

A

Polyacrylamide Gel Electrophoresis (SDS-PAGE)

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10
Q

Charge imparted during SDS-PAGE process on proteins

A

Negative

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11
Q

Gel used SDS-PAGE is made of

A

Gel is made of Polyacrylamide and protein mixture is loaded on top

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12
Q

Energy in 1gm of carbs, proteins and fats?

A

4, 4, 9 calories

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13
Q

Five carbon aldoses in order

A

Ribose, Arabinose, Xylose and Lyxose

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14
Q

Six carbon aldoses in order

A

Allose, Altose, Glucose, Mannose, Gulose, Idose, Galactose and Talose

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15
Q

Five carbon Ketoses in order

A

Ribulose and Xylulose

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16
Q

Six carbon ketoses in order

A

Psicose, Fructose, Sorbose and Tagatose

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17
Q

Monosaccharides in maltose

A

alpha glucose and alpha glucose

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18
Q

Monosaccharides in Lactose

A

beta Galactose(1) and alpha Glucose(4)

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19
Q

Monosaccharides in Sucrose

A

beta fructose(2) to alpha glucose(1)

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20
Q

Monosaccharides in Trehalose

A

alpha Glucose(1) to Alpha Glucose(1)

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21
Q

The branching in amylopectin is of the type

A

(alpha1 to alpha6)

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22
Q

Role of Dextran

A

extracellular adhesive in baceteria

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23
Q

Role of Peptidoglycan

A

gives rigidity and strength to cell envelope in bacteria

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24
Q

Significance of Hyaluronan

A

Extracellular matrix of skin and connective tissue, viscosity and lubrication of joints in vertebrates

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25
Q

What are phospho-sphingolipids made of?

A

Sphingoline, Fatty acid and (PO4 - Choline)

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26
Q

What are GlycoSphingolipids made of?

A

Sphingoline, Fatty acid and Mono- or Oligo- saccharide

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27
Q

What are Galactolipids?

A

Galactolipids are Glycolipids made of Glycerol, 2 Fatty Acids and a (Mono- or Oligo- Saccharide - SO4)

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28
Q

What are Archeal ether lipids made of?

A

(Glycerol - PO4) connected to another glycerol by two Diphytanyl ether linkages which itself is connected to another Glycerol by a PO4

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29
Q

What are simple triacylglycerols?

A

Glycerols with the same fatty acids at all 3 positions

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30
Q

The three simple tryacylglycerols 16:0, 18:0 and 18:1 are

A

tripalmitin, tristearin and triolein

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31
Q

What is fat tissue?

A

Adipocytes

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32
Q

What is a primer?

A

Primer is a short nucleic acid sequence that provides a starting point for DNA synthesis

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33
Q

What are omega three fatty acids?

A

Poly Unsaturated Fatty Acids with a double bond between C-3 and C-4 from the methyl side are called omega 3 fatty acids

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34
Q

Why are omega-3 PUFAs important

A

A dietary ratio of 1:1 to 1:4 of omega-3 and omega-6 should be maintained to avoid cardiovascular disease. ratio in diet in average north american is 1:10 to 1:30

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35
Q

At room temperature saturated fatty acids 12:0 to 24:0 and unsaturated fatty acids are

A

waxy solids and liquids

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36
Q

What is good cholesterol and bad cholesterol?

A

LDL is bad🤢 and HDL is good (trans fat increases LDL)

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37
Q

How many types of membrane lipids are there and what are they?

A

5 and glycerophospholipids, (galactolipids and sulfolipids), tetraether lipids, sphingolipids and sterols

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38
Q

Glycerophospholipids structure

A

two fatty acids joined to glycerol

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39
Q

galactolipids and sulfolipids structure

A

two fatty acids esterified to glycerol, but lack the phosphate of phospholipids.

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40
Q

tetraether lipids

A

two very long alkyl chains are ether-linked to glycerols at both ends

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41
Q

sphingolipids structure

A

a single fatty acid is joined to fatty amine, sphingosine

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42
Q

sterols

A

rigid system of four fused hydrocarbon rings

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43
Q

What type of lipid determines blood group?

A

Glycosphingolipid

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44
Q

What is the oligosaccharide in the lipid that determines the bloodgroup that gives O,A.B antigen?

A 
O Antigen (Glu - Gal - GlcNAc - Gal) - Fuc
A Antigen (Glu -Gal - GlcNAc - Gal)( - Fuc)( - GalNAc)
B Antigen (Glu -Gal - GlcNAc - Gal)( - Fuc)( - Gal)
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45
Q

What is the primary structure of a protein?

A

the linear sequence of amino acids: Gly-Ala-Lys-Trp

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46
Q

What is the secondary order of proteins?

A

stable arrangement of amino acids giving rise to recurring structural patterns, such as α-helix and β-sheet.

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47
Q

What secondary structure is present in myoglobin?

A

alpha helix

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48
Q

Tertiary structure of proteins

A

three-dimensional folding of a polypeptide chain.ular.

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49
Q

Quaternary structure of proteins

A

when a protein has two or more polypeptide chains, their arrangement in space.

50
Q

Groups other than amino acids are called

A

Prosthetic Groups

51
Q

protein with carb prosthetic group

A

Immunoglobin G

52
Q

protein with phosphate prosthetic group

A

Casein of Milk

53
Q

protein with flavo nucleotide prosthetic group

A

Succinate dehydrogenase

54
Q

protein with zinc prosthetic group is

A

Alcohol dehydrogenase

55
Q

protein with calcium pros group

A

Calmodulin

56
Q

protein with molybdenum pros group

A

dinitrogenase

57
Q

protein with copper pros group

A

Plastocyanin

58
Q

components of nucleotide

A

Nitrogenous Base, Pentose Sugar and one or more phosphates

59
Q

Nucletide without phosphate group is called

A

Nucleoside

60
Q

DNA contains which pentose sugar

A

2’-deoxy-D-ribose

61
Q

the pentose sugar in RNA is

A

D-ribose

62
Q

Number of hydrogen bonds in adenine-thymine bond

A

two

63
Q

number of hydrogen bonds in Guanine-cytosine bond

A

3

64
Q

Erwin Chargaff’s Rules are

A

1) The base composition of DNA generally varies from one species to another.
2) DNA specimens isolated from different tissues of the same species have the same base composition.
3) The base composition of DNA in a given species does not change with an organisms’s age, nutritional status, or changing environment.
4) In all cellular DNA, A=T and G=C. (A+G = T+C)

65
Q

How far are vertically stacked bases in DNA?

A

3.4A

66
Q

How are the strands in DNA aligned?

A

Anti-Parallel

67
Q

What are the 3 forms of DNA and which is the most stable form?

A

A, B and Z. B being the most stable

68
Q

What does RNA do in DNA synthesis and what types of RNA are involved?

A

RNA carries genetic information from DNA to the protein biosynthetic machinery of the ribosome.

By the process of transcription, RNA is synthesized from the DNA template.

Three types of RNAs involved in protein synthesis:
mRNA: Messenger RNA
tRNA: Transfer RNA
rRNA: Ribosomal RNA

69
Q

Tertiary Structure of RNA

A

Single Strands
Bulge
Internal loop
Hairpin

70
Q

What are the three types of cloning?

A

Reproductive cloning/organism cloning (legally restricted)
Therapeutic cloning/stem cell cloning
Molecular cloning/DNA cloning

71
Q

Removal of nucleus from the egg is called

A

Enucleation

72
Q

Circular piece of DNA used in cloning is called?

A

Plasmid

73
Q

What is a plasmid?

A

A plasmid is a small, extrachromosomal DNA within a cell that is physically separated from chromosomal DNA and can replicate independently.

74
Q

What is the full form of REs?

A

RE = Restriction Endonucleases

75
Q

How many types of REs are there and what are they?

A
  1. Type 1 REs cleave at random sites
    Type 3 REs cleave DNA about 25bp from recognition site
    Type 2 REs cleave DNA with recognition site itself. They dont require ATP. They also make staggered or cleaved ends
76
Q

What are the ways to introduce plasmids into bacterial cells?

A 
Transformation:
The cells (often E. coli) and plasmid DNA are incubated together at 0 C in a calcium chloride solution, then subjected to heat shock by rapid shifting the temperature to between 37 C and 43 C.
Electroporation:
The cells (often E. coli) are incubated with plasmid DNA are subjected to high-voltage electric pulse. This approach transiently renders the bacterial membrane permeable to large molecules.
77
Q

What is ori?

A

Origin of replication (ori), a sequence where replication is initiated by
cellular enzymes

78
Q

Size of plasmids ranges from?

A

5000 to 400000 bp

79
Q

Flourescence has a lower or higher wavelength than absorbed light?

A

Higher obv🤷‍♀️

80
Q

What is the difference between the excitation and emission peaks called?

A

Stokes shift

81
Q

Describe protein labelling strategies

A

1) Genetic encoding: C-terminal or N-terminal
pros: specificity
cons: time taking
2) Antibody based detection
pros: quick
cons: not suitable for live cells
non specific
3)Covalent modification
1) Using Self-labelling enzyme derivatives
2) Using chemical labelling
pros: easy and efficient
cons: suitable for in-vitro studies only
3)Enzymatic labelling:
Fast Efficient and selective for both in-vivo and vitro targetting whole proteins or cells

82
Q

Where was GFP first discovered?

A

In the jellyfish Aequorea victoria

83
Q

What causes the fluorescence of GFP?

A

Fluorescence is due to amino acids at positions S65. T66 and G67

84
Q

What is quantum yield?

A

Ratio of number of photons absorbed to number of photons emitted

85
Q

Disadvantages of fluorescent proteins over dyes

A

Use of FPs requires genetic engineering, a time consuming process
Fusion of protein of interest with FP increases its mol. weight by ~27 kD, which may affect protein function.
Fast photobleaching compared to fluorescent dyes
Limited spectral range compared to fluorescent dyes
Less brightness compared to fluorescent dyes
Most FPs tend to oligomerize

86
Q

What is the first fluorescent dye invented?

A

fluorescein

87
Q

Fluorescence recovery after photobleaching (FRAP)

A

Used to study protein mobility in cells and diffusion dynamics

88
Q

What is the formula for the diffusion coefficient

A

D = 0.224\frac{r^2_n}{\tau^\frac{1}{2}}

89
Q

What is mobile fraction?

A

Mobile fraction: fraction of fluorescent protein diffusing into bleached region during the time course of the experiment.

90
Q

What is immobile fraction?

A

Immobile fraction: fraction of immobile bleached molecules within the bleach spot that don’t allow fluorescent molecules to occupy their space.

91
Q

Efficiency of FRET based on distance is

A

inversely proportional to sixth power of dsitance

91
Q

Efficiency of FRET based on distance is

A

inversely proportional to sixth power of distance

92
Q

What is FRET

A

Forster or Fluorescence Resonance Energy Transfer (FRET) is a mechanism describing energy transfer between two light-sensitive molecules (chromophores).
FRET is used to check protein-protein, protein-DNA interactions and protein conformation dynamics.

93
Q

What are optical tweezers?

A

Optical tweezers use tightly focussed light beam (IR laser) to hold and manipulate microscopically small objects.

94
Q

What are optical tweezers?

A

Optical tweezers use tightly focussed light beam (IR laser) to hold and manipulate microscopically small objects.

95
Q

What does DNA ligase do and how?

A

DNA ligase is used to join two DNA molecules. It forms phosphodiester bonds between the DNA molecules.
A vector digested by an RE can only be ligated to insert digested by same RE

96
Q

Features of BACs?

A

They can clone larger DNA fragments
They have stable ori that limits the number of plasmids in a cell
BACs contain a selection and screening marker

97
Q

What is LacZ and what does it do?

A

LacZ stands for beta-galactosidase which converts X-gal(white) to a blue product. If LacZ is intact colony will be blue otherwise will be white

98
Q

Plasmids usable in more than 2 species is called

A

Shuttle vector

99
Q

Elements of YAC

A

yeast Ori, two selectable markers, and specialized sequences (derived from centromeres and telomeres) needed for stability and proper segregation of chromosomes at cell division.

100
Q

What are expression vectors?

A

Cloning vectors with the transcription and translation signals needed for the regulated expression of a cloned gene are called expression vectors.

101
Q

What reduces mobile fraction?

A

Aggregation of proteins, binding and diffusional barries reduce mobile fraction

102
Q

Stages of DNA packing

A

DNA > Nucleosomes > Chromatine > Chromatine Loops > Condensed Chromatine Loops > Chromosome

103
Q

Phases of mitosis are

A

Interphase, Prophase,Mrtaphase,Anaphase and Telophase

104
Q

Is chromosome organization non-territorial or territorial?

A

Territorial

105
Q

Steps of laser-UV-microirradiation

A

1) The nucleus of the live cell was microirradiated in G1
2) Pulse-labeled with 3H thymidine
3) Fixed immediately
4) Autoradiography

106
Q

What are integral proteins of the nuclear envelope?

A

Lamina and emerin. They are linked to rare genetic disorders called nuclear envelopathies.

107
Q

Give examples of envelopathies

A

Hutchinson-GIlford progeria syndrome and Emery-Dreifuss muscular dystrophy caused by repositioning of chromosomes 13 and 18

108
Q

Steps of FISH

A

Fix > Permeablize > Denaturation > Hybridization > Wash to remove unbound and non-specific probes > DAPi > Coverslip

109
Q

Types of probes for DNA FISh

A

Centromere probes
Whole Chromosome
Telomere
Locus

110
Q

What does oxidoreductase enzyme do?

A

Transfer of electrons

111
Q

Transferase enzyme

A

Group Transfer reaction

112
Q

Hydrolases

A

Hydrolysis reactions

113
Q

Lyases

A

Cleavage of C-C,C-O,C-N

114
Q

Isomerases enzymes

A

Yields isomeric forms

115
Q

Ligases

A

Formation of C-C,C-S,C-) and C-N by cleavage of ATP

116
Q

Relation between Substrate Concentration [S] and Reaction Rate[V_0]

A

V_0 = \frac{V_{max}[S]}{K_m + [S]}

where K_m is the Michealis constant

117
Q

What is the michealis Menten equation

A

V_0 = (V_max[S])/(K_m + [S])

118
Q

What is the Lineweaker Burk Equation

A

1/V_0 = K_m/(V_max[S]) + 1/V_max

119
Q

Specificity Constant si

A

K_cat/K_m

120
Q

Unit of enzyme activity is

A

katal = 1micromole/min = 16.67nanomole/s

Life Sciences (1 decks)

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