Biotechnology

Question 1

Many bacteria and certain fungi contain self-replicating extrachomosomal DNA molecules called

Answer 1

Plasmids

Question 2

Plasmid DNA is

Answer 2

Closed circular and double-stranded

Question 3

Self-replicating and do not integrate into the host genome

Answer 3

Plasmids

Question 4

The smallest plasmids

Answer 4

contain only a few thousand base pairs

Question 5

The largest plasmids

Answer 5

contain over 100,000 base pairs

Question 6

Confer a specialized and sometimes protective characteristic to the organism

Answer 6

Genetic information for the translation of proteins found in bacterial plasmids

Question 7

Certain plasmids carry the gene for the enzyme

Answer 7

Beta-lactamase

Question 8

Catalyzes the decomposition of certain antibiotics such as penicillin and amoxicillin

Answer 8

Beta-lactamase

Question 9

Able to pass from an antibiotic-resistant cell to an antibiotic sensitive cell of the same or another bacterial species, thus rendering the latter resistant

Answer 9

Plasmids

Question 10

Posess an ampicillin resistance gene and a Beta-lactamase gene

Answer 10

A plasmid from the strain of Escerichia coli (JM101)

Question 11

One of the fastest methods to isolate plasmid DNA

Answer 11

Single-use columns

Question 12

Based on the principle of anion exchange chromatography

Answer 12

Single-use columns

Question 13

DNA binds to a modified

Answer 13

Silica matrix and is eluted using sterile distilled water

Question 14

Isolation of Plasmid DNA Materials

Answer 14

1. Pre-Lysis Buffer
2. Alkaline Lysis Solution
3. Neutralizing Solution
4. Glassmilk Spinbuffer
5. Wash Solution
6. Sterile Distilled Water
7. RPM Spin Filters
8. Catch Tube

Question 15

To resuspend the cell pellet

Answer 15

Add 50 micro liters Pre-Lysis Solution and mix by vortexing or pipetting up and down until completely suspended

Question 16

A white precipitate will form consisting of

Answer 16

Cell membranes, proteins, and chromosomal DNA

Question 17

To elute plasmid DNA from the Spin Filter,

Answer 17

Add 50 micro liters of sterile water and spin for 2 minutes to collect DNA in the bottom of the vial

Question 18

Many types of bacterial can transfer ________ between individual cells.

Answer 18

DNA

Question 19

Under certain conditions, large portions of chromosomes are transferred to the recipient cell

Answer 19

transferred to the recipient cell

Question 20

Under some condition smaller autonomously replicating circular DNA molecules called _______ are transferred

Answer 20

Plasmids

Question 21

Plasmid DNAs do not usually

Answer 21

encode information essential for the functioning of the cell

Question 22

Bacteria that lack plasmids

Answer 22

function normally

Question 23

Bacteria that possess plasmids

Answer 23

may be capable of performing additional functions, as for example showing resistance to specific antibodies

Question 24

Some bacteria display _____________, while others do not

Answer 24

Competence

Question 25

Ability to take up DNA at a particular stage in the life cycle

Answer 25

Competence

Question 26

It is possible to make E. coli cells competent by treatment with

Answer 26

certain reagents

Question 27

When the introduction of foreign DNA into a cell alters the phenotype of the cell, the cell is said to have been

Answer 27

Transformed

Question 28

The process of introducing DNA into a cell is known as

Answer 28

Transformation

Question 29

Most procedures for the transformation of E. coli are based on

Answer 29

The observation that when treated with cold CaCl2 and briefly heat-treated, bacteria could be transformed

Question 30

How CaCL2 confers competence to E. Coli

Answer 30

Is unknown, as is the mechanism by which DNA enters the cell

Question 31

A number of factors influence the efficiency of transformation including:

Answer 31

1. The strain of bacteria 2. The growth medium 3. The stage of the cell cycle

Question 32

In order to detect cells that have taken up plasmid DNA, it is necessary for the plasmid to contain a

Answer 32

Marker

Question 33

A gene that when expressed as RNA or protein, reveals the presence of that gene

Answer 33

Marker

Question 34

Frequently markers include ________ that confer resistance to antibiotics to which the cell is ordinarily sensitive

Answer 34

genes

Question 35

Purpose of the Transformation of Plasmid lab

Answer 35

To demonstrate phenotype changes in bacteria that have been transformed with an antibiotic-resistance gene and a metabolic marker

Question 36

E. coli cells are transformed with

Answer 36

The plasmid pBLU

Question 37

Carries genes for two identifiable phenotypes: Amp and lacZ

Answer 37

Plasmid pBLU

Question 38

Two identifiable phenotypes

Answer 38

Amp and lacZ

Question 39

Responsible for bacterial resistance to Beta-lactam antibiotics

Answer 39

Amp gene

Question 40

Codes for Beta-lactamase

Answer 40

Amp gene

Question 41

Expresses resistance to the antibiotic ampicillin

Answer 41

Beta-lacatamase

Question 42

Widely used reporter gene which cods for Beta-galactosidase

Answer 42

lacZ

Question 43

5-bromo-4-chloro-3-indolyl-Beta-galactopyranoside

Answer 43

X-gal (lactose analog broken down by the enzyme)

Question 44

Produce a visible blue product

Answer 44

The enzyme breaks down the lactose analog X-gal

Question 45

The recipient E. coli strain lacks

Answer 45

amp and lacZ (thus transformed cells are converted to an amp and lacZ phenotypes

Question 46

Selective agent

Answer 46

Ampicillin

Question 47

Only _________ cells are able to grow on medium containing amipicllin

Answer 47

transformed

Question 48

Not a selective marker, but only cells transformed with the plasmid metabolize _________ to produce blue colonies

Answer 48

X-gal

Question 49

Luria-Bertania Media

Answer 49

``` Deionized Water Bactotryptone Bacto yeast NaCL 5M NaOH 1M HCL ```