MCM Final - Tools of Cell and Molecular Biology

Question 0

fluorescence microscopy

Answer 0

use fluorescent tags to visualize a molecule

-absorb light at one wavelength, emit at another

-

Question 1

light microscopy

Answer 1

use light to visualize cells and tissues

Question 2

in vitro

Answer 2

in glass

Question 3

in vivo

Answer 3

in the living

Question 4

medium for cell culture

Answer 4

salts, carbohydrates, amino acids, and vitamins

supplemented by animal serum
-contains growth factors

Question 5

passaging

Answer 5

process of removing cells from a surface and replating at a lower density

Question 6

explants

Answer 6

culture a small tissue fragment

-usually in neuroscience research

Question 7

primary cultures

Answer 7

prepared directly from tissues of an organism

can be homogenous or heterogenous

don’t last very long

Question 8

immortalized cell lines

Answer 8

over come replicative cell senescence

generated from cancer cells

created by introducing oncogenes

Question 9

Hayflick Limit

Answer 9

human somatic cells have a limited number of divisions (50-60)

Question 10

first line of immortalized cells?

Answer 10

HeLa cell line from Henrietta Lacks

Question 11

hybridoma cell line

Answer 11

used to produce monoclonal antibodies

- infinite quantity of identical antibodies
- used in diagnosis and treatment

Question 12

heterocaryon

Answer 12

cell with two nuclei

Question 13

process to create hybridoma cell line?

Answer 13

1 create hybrid cell type
-from differentiated normal cell and immortalized cell
2 cells fuse and create a heterokaryon
-use b lymphocytes from mouse and b lymphocyte tumor cells
3 only hybridoma cells survive in the selective medium
4 cells multiply and scan for anti-X antobidy
5 positive clones are a continuous source of anti-X antibody!

Question 14

antibody structure and function

Answer 14

y shaped
2 Fab domains - antigen binding
1 Fc domain - signaling

one lymphocyte creates one identical antibody in large quantities

Question 15

polyclonal antibodies

Answer 15

antibodies produced by inoculating an animal against an antigen
-from multiple lymphocytes agains multiple epitopes

Question 16

Fab Domain

Answer 16

2 on antibody

-allow the binding of antigen

Question 17

Fc Domain

Answer 17

1 on antibody - allow signaling to occur

Question 18

primary antibody

Answer 18

bind antigen

Question 19

secondary antibody

Answer 19

bind Fc portion of primary antibody (bound to antigen)

has a marker than can be identified

Question 20

immunocytochemistry

Answer 20

stain cells

Question 21

immunohistochemistry

Answer 21

stain tissues

Question 22

enzyme-linked immunosorbent assay (ELISA)

Answer 22

marker in enzyme that creates a color precipitate when binds substrate
-used in sensitive tests (infection, pregnancy)

Question 23

electrophoresis of DNA and mRNA

Answer 23

negative charged phosphate groups

movement based on size of fragment

Question 24

SDS

Answer 24

covers protein during electrophoresis to give it an overall negative charge
-so it can be electrophoresed

also, need to unfold protein before electrophoresis

Question 25

Northern Blotting

Answer 25

mRNA detected with labeled oligonucleotide probe

Question 26

Southern Blotting

Answer 26

DNA sequence detected with a labeled oligonucleotide probe

Question 27

Western Blotting

Answer 27

aka immunoblotting

-protein detected using primary and secondary antibodies

Question 28

recombinant DNA

Answer 28

method of producing large quantities of DNA, RNA, or protein
-molecular cloning

source DNA - fragment isolated using RFLPs (restriction endonuclease sites)

Question 29

cDNA

Answer 29

copy DNA
-need to remove introns if protein synthesized in bacteria

-make cDNA from mRNA using reverse transcriptase

Question 30

process of creating cDNA

Answer 30

1 make cDNA from mRNA using reverse transcriptase
2 add oligonucleotide linkers with restriction endonuclease cleave sites to the ends of cDNA
3 cDNA ligated to appropriate vector

Question 31

plasmid vector

Answer 31

DNA molecule capable of independent replication when placed in host cell

- contain:
	- restriction endonuclease site (same as source DNA)
	- gene for antibiotic resistance
	- origin of replication

Question 32

recombinant vector

Answer 32

combination of source DNA and vector DNA

Question 33

recombinant proteins

Answer 33

produce large amounts of protein for therapeutic use

Question 34

expression vectors

Answer 34

in recombinant protein

• require DNA sequences that express protein
  
  • include promoter and start site

Question 35

gene therapy

Answer 35

introducing normal copy of gene into a cell

Question 36

retrovirus

Answer 36

carry RNA genome

- reverse transcribe it into DNA
- DNA inserted randomly into host DNA
- requires host cell to be actively dividing
- permanent change

Question 37

adenovirus

Answer 37

carry DNA genome

- not integrated into host DNA
	- eventually lost
- host cells do not need to be actively dividing

Question 38

transgenic animals

Answer 38

inject cDNA into fertilized ova

if successful - offspring are chimeric in somatic and stem cells
-next generation will not be