Biology Of The Cell Lab Exam 2

Front 1

what is the Central Dogma of Molecular Biology?

Back 1

DNA:Replication -> DNA:Transcription -> RNA:Translation ->
Protein

Front 2

What is a gene?

Back 2

a nucleotide sequence in DNA to which a specific genetic function can be assigned.

Front 3

mRNA

Back 3

messenger RNA

Front 4

what does messenger RNA do?

Back 4

takes its genetic information or message out of the nucleus and into the cytoplasm of the cell.

Front 5

converts the nucleotide sequence of the mRNA into a specific sequence of amino acids to produce a specific protein.

Back 5

translation

Front 6

translation of mRNA is accomplished by translating a series of three nucleotides

Back 6

codon

Front 7

what molecules are involved in the central dogma of molecular biology?

Back 7

DNA RNA Proteins

Front 8

what are the processes involved in the central dogma of molecular biology?

Back 8

Replication, Transcription, translation

Front 9

what is a peptide bond?

Back 9

chemical bond between the carbonyl group of one amino acid and the amio group of a second amino acid (a special form of amide linkage)

Front 10

what are the start codons?

Back 10

met (AUG)

Front 11

what is the stop codons?

Back 11

UAA, UAG, UGA

Front 12

what amino acid is the start codon?

Back 12

Met

Front 13

what are restriction endonucleases?

Back 13

they are enzymes that catalyze the cleavage of the phosphdiester bonds within both strands of DNA.

Front 14

nucleotide sequence that is identical to its complementary strand when each is read in the same chemical direction

Back 14

palindromic sequences

Front 15

how are palindromes related to restriction enzymes?

Back 15

restricted enzymes only cut at very specific palindronic sequences of bases.

Front 16

what are "sticky" or "cohesive" ends?

Back 16

staggered cleavage site that can rejoin again.

Front 17

what are "blunt" ends?

Back 17

cuts that are opposite of each other.

Front 18

what is a recognitioon site where there are only four possible sequences?

Back 18

degenerate

Front 19

what are the recognition sites that are divided by a certain number of totally variable bases?

Back 19

hyphenated sites

Front 20

a powerful separation method frequently used to analyze DNA fragments generated by restriction enzymes

Back 20

agarose gel electrophoresis

Front 21

what does the gel consists of in a gel electrphoresis?

Back 21

microscopic pores that act as a molecular sieve.

Front 22

Since DNA has a strong _____ charge at neutral pH, it migrates through the gel towards the ____ electrode during electrophoresis.

Back 22

negative; postive

Front 23

the ____ the lnear fragment the faster it migrates in gel electrophoresis.

Back 23

smaller

Front 24

what was our source of DNA in gel electrophoresis?

Back 24

Lambda DNA

Front 25

Lambda phage DNA contains 10-16 base single-stranded regions at the 5'-3' terminus that are self-complimentary called___.

Back 25

cos ends

Front 26

what are our source of emzyme in gel electrophoresis? (6)

Back 26

Hind III, Hae III, Eco RI, Eco RII, Bgl I, Bam HI

Front 27

What is the restriction enzyme?
Bacillus globigii,
Bacillus amyloliquefaciens H

Back 27

Bgl I; Bam HI

Front 28

What is the restriction enzyme?
Escherichia coli strain RY 13,
Escherichia coli strain R 245

Back 28

Eco RI; Eco RII

Front 29

What is the restriction enzyme?
Haemophilus segyptius,
Haemophilus influenzae Rd

Back 29

Hae III, Hind III

Front 30

what is the horizontal axis in a restriction digest plot?

Back 30

migration distance

Front 31

what is the vertical axis in a restriction digest plot?

Back 31

log bases pairs

Front 32

how many amino acids are there?

Back 32

20

Front 33

the sequence or order of amino acids along a polupeptide chain is referred to as the ____ of the protein.

Back 33

primary structure.

Front 34

the spatial arrangement of the protein backbone that is generated from the folding of the polypeptide chian is called the _____ of the protein.

Back 34

secondary structure

Front 35

what is a central organizing feature of enzymes, antibodies, and most othe proteins?

Back 35

B-sheet

Front 36

the ____ of a potein describes the detailed features of the three dimensional conformation of the polypeptide chain.

Back 36

tertiay structure

Front 37

the ____ of a protein is due to the type and sequence of its constituent amino acids.

Back 37

three-dimensional structure

Front 38

the unfolding of a proteins conformation

Back 38

denauturation

Front 39

the movement of charges molecule under the influence of an electric feild.

Back 39

electrophoresis

Front 40

what are the difference between protein electrophoresis and DNA electrophoresis?

Back 40

DNA is (-) so it migrates to the (+) end of the gel. Amino Acids in Proteins are either (+) or (-) so they move in both directions.

Front 41

what is an iso-electric point?

Back 41

the pH at which an amino acid or protein does not migrated in an electric feild

Front 42

what is the role of pH of electrophoresis buffer n selarating proteins out of solution?

Back 42

it detemines the net charge on the protein molecules in the sample

Front 43

what will happen if you increase or decrease the pH of the buffer?

Back 43

the higher the charge the faster the protein will travel. so if the protein's pH is close to the pH buffer it will move slower. if the pH of the protein is farther from the pH buffer it will move faster.

Front 44

what type of buffer was used in protein electrophoresis?

Back 44

electrophoresis buffer

Front 45

what is genetic transformation?

Back 45

change caused by genes and it involves the insertion of a gene(s) into an organism in order to change the orgamism's traits

Front 46

who normally do genetic transformation?

Back 46

agriculture, bio-remediation, and medicine

Front 47

what is the procedure of genetic transformation?

Back 47

reqires two steps:
1. a calcium cholorid buffer which offsets the environment to neutralize the similar charges
2. heat shock which increase the permeability of the bacterial membrane

Front 48

what is your vessel for ntroducing genes?

Back 48

plasmid (pGLO)

Front 49

what must be done to your organism to get this vessel in?

Back 49

heat shock to break bacterial membrane

Front 50

provides a solid matrix to support bacterial growth

Back 50

agar

Front 51

a carbodrate, normally used as source of food by bacteria

Back 51

arabinose

Front 52

a protein the provides resistance to the antibiotic amplicillin

Back 52

beta-lactamase

Front 53

appluing bilogy in the real world by the specific maipulation of living organisma to produce potentially beneficial products

Back 53

biotechnology

Front 54

when a population of cells is prepared by growth from a single cell, all the cells in the poulation will be geneticlly identical

Back 54

cloning

Front 55

a clump of genetically identical acterial cells groowing on an agar plate.

Back 55

colony

Front 56

the liquid and solid media are referred to as LB broth and agar are made from an extract of yeast and an enzymatic digest of meat byproducts the provides a misture wich are nutrients for bacterial growth.

Back 56

culture media

Front 57

the manilulation of an oganism's genetic material by introducing or eleminating specific genes.

Back 57

genetic engineering

Front 58

polymerizes when heated to form a solid gel and functons to provide a solid support on which to culture the bacteria

Back 58

agar

Front 59

the unique 3-D conformation cuases it to resonate when exposed to ultraviolet and give off energy in the form of visible green light.

Back 59

green fluorescent protein

Front 60

a circular DNA molecule, capable of autonomus replication, carrying one or more genes for antibiotic resistance proteins and a cloned foreign gene such as GFP

Back 60

plasmid

Front 61

plasmid containing the GFP sequence and ampicillin resistance gene which codes for beta-lactamase

Back 61

pGLO

Front 62

the process of cutting and recombining DNA fragments as a means to isolate genes or to alter their structure and function

Back 62

recombinant DNA technology

Front 63

process of iddentifying wanted bacteria from a bacterial library

Back 63

screening

Front 64

minimizing the possibility of outside bacterial contamination during an experiment through observance of cleanliness and using careful labroatory techniques

Back 64

sterile technique

Front 65

process of passing an inoculation loop with bacteria on it across an agar plate

Back 65

streaking

Front 66

an autonomously replicating DNA molecule into which foreign DNA fragments are inserted and then paropagated in a host cell

Back 66

vector

Front 67

what plasma did we use for transformation lab

Back 67

pGLO

Front 68

what genes did pGLO contain?

Back 68

GFP and a gene for resistance to the antibiotic ampicillin.

Front 69

why did we use E. coli in transformation lab?

Back 69

Because it is a bacteria that reproduces fast.

Front 70

what specific E. coli did we use in transformation lab and why?

Back 70

E. coli HB 101 because it is non-pathogenic, non-infectious, and only grows in the presence of cetain media

Front 71

what is a phenotype?

Back 71

the observable character of a cell or organism

Front 72

what is amp?

Back 72

ampicillin: we put ampicillin into the plates to kill off the bacteria that doesnt have the GFP gene or the antibiotic gene. (if doesnt have antibiotic gene then doesnt have GFP gene so we dont need them)

Front 73

what is LB?

Back 73

luria-bertani: which is the liquid or solid media where the nutrients neeed to make bacteria grow is located

Front 74

what is ara?

Back 74

arabinose: which turns on the GFP in transformed cells

Front 75

the liquid that is present above the pellet is called____

Back 75

supernatant

Front 76

HIC

Back 76

hydrophobic interaction chromatography

Front 77

when the lasate is applied to the column, the hydrophoi proteins that are applied the column in a high salt buffer will stick to the beads while all other proteins in the mixture will pass through. when the salt is decrease, the hyfrophobic proteins will no longer stick to the beased and will drop out the botton of the column in a purified form

Back 77

HIC

Front 78

spinning a mixture at very high speed to separate heavy and light particles

Back 78

centrifuge

Front 79

what is the result of centrifuge?

Back 79

a pellet and supernatant

Front 80

what did the pellet in the expression lab contain?

Back 80

bacteria

Front 81

what happens to GFP in each buffer? (Binding, Wash, and Elution)

Back 81

Binding: the GTP changes its comfomation to where the hydrophobic parts are on the outside sticking to the beads.
Wash: the weaker hydrophobic parts drop through the column
Elution: the GFP returns to its original conformation and drops through the column.

Front 82

enzyme needed to break open bacteria cell walls

Back 82

lysozyme

Front 83

a process for separating complex liquid mistures of proteins or other molecules by passing a liquid misture over a column containing a solid mixture.

Back 83

chromatography