Quiz 3 - Pt. 1 How Cellular Information is Altered

Question 1

We can alter cells by using ____ or genetic engineering. ____ is subjecting the cells to stress causing changes in the genetic make-up.

Answer 1

mutation

Question 2

________ is the purposeful transfer of DNA from one type of organism to another.

Answer 2

Genetic Engineering`

Question 3

mistakes in the genetic code (can arise from replication and/or damage)

Answer 3

Mutations

Question 4

organism with a genetic mutation

Answer 4

Mutant

Question 5

the organism without the genetic organism

Answer 5

Wild type

Question 6

genetic construction of an organism

Answer 6

Genotype

Question 7

characteristics expressed by an organisms.

Answer 7

Phenotype

Question 8

usually refers to transcription+translation+posttranslation processing

Answer 8

Expression

Question 9

EXAMPLE:

• Strain A has the tol operon for toluene degradation, and is in a reactor growing on glucose.
• Strain B has the tol operon for toluene degradation, and is in a reactor growing on toluene.
• These strains have the same _____, but ________

Answer 9

genotype
different phenotypes

Question 10

base change may or may not result in an amino acid change

Answer 10

Consequences

Question 11

If the amino acid is the same as before the mutation there is ________.

Answer 11

no consequence

Question 12

If the amino acid is different, but not in the region of the ______, there may be no consequences

Answer 12

active site

Question 13

If the mutation is in the active site, there may be some _______________

Answer 13

enzyme activity consequence

Question 14

If the mutation changes the amino acid to a stop codon, the resulting protein will be _______ and probably ________.

Answer 14

truncated (shortened) and probably not active

Question 15

confers upon the mutant an advantage for growth, survival or detection under a set of environmental conditions that the wild type does not have.

Answer 15

Selectable mutation

Question 16

Examples of Selectable Mutation or Selection

Answer 16

• Antibiotic resistance
• Ability to grow on toluene
• Inability to produce lysine
• Ability to produce bioluminescence
• Ability to produce more of an enzyme
• Inability to grow at higher temperatures

Question 17

Natural Mutation Rates
* 10^-3 - 10^-9 =
* 10^-6 =

Answer 17

mutations per cell conversion
1 mutation/1,000,000 divisions

Question 18

Increase Mutation Rates
____ : chemicals, radiation
_________ (i.e. lots of divisions)

Answer 18

Mutagens
Lots of growth

Question 19

Why do we want to increase mutations?

• We want a cell to develop ____________ that are _______ for us.
• For example, ________ inhibition of lysine to increase lysine ______

Answer 19

• specific characteristics, advantages
• removing feed back
• production

Question 20

uptake of free DNA by a cell. The cell membrane has to be permeable to DNA.

Answer 20

Transformation

Question 21

DNA is carried into the call in a phage.

Answer 21

Transduction

Question 22

Cell to cell transfer of DNA. Also called mating.

Answer 22

Conjugation

Question 23

Natural Gene Transfer/Rearrangement:

Once the DNA is inside the cell it can remain ______ from the chromosome in self-replicating _________ or _____ into the chromosome.

To _____, the DNA must be complementary to the _________________ on the ends.

Answer 23

separate
self replicating plasmid
Integrate

integrate
chromosomal DNA

Question 24

Using _________ engineers and microbiologists were able to increase penicillin from 0.001 g/L to 50 g/L

Answer 24

mutation and selection

Question 25

Genetic Engineering: Using _____ to **purposefully manipulate DNA**. The DNA is ______ **outside** of the cell, and then sent into the cell

Answer 25

natural mechanisms manipulated

Question 26

Genetic Engineering Tools

Answer 26

Restriction enzymes Gel electrophoresis (Southern Blot) Polymerase Chain Reaction (PCR) Plasmid

Question 27

enzymes that **cut DNA** at specific sequences. Different enzymes will cut at different sequences.

Answer 27

Restriction enzymes

Question 28

A method to detect what **sizes of DNA** a sample contains.

Answer 28

Gel electrophoresis (Southern Blot)

Question 29

A process used to make many **copies** of a piece of **DNA**.

Answer 29

Polymerase chain reaction (PCR)

Question 30

**self replicating**, **circular piece of DNA** that can survive in a cell.

Answer 30

Plasmid

Question 31

DNA is denatured by ______ Renaturation on ______

Answer 31

heating cooling

Question 32

The ____ cuts both **DNA strands** at the same site. DNA fragments join at ______ And Formed a ________

Answer 32

enzyme sticky ends Recombinant DNA

Question 33

Organism from which derived: 1. Anabaena Variabilis 2. Bacillus amyloliquefaciens 3. Bacillus globigii 4. Escherichia coli RY 13 5. Escherichia coli R245 6. Haemophilus aegyptius 7. Haemophilus haemolyticus 8. Haemophilus inflenzae RD 9. Haemophilus parainflanzae What are the Enzyme called?

Answer 33

Enzyme: 1. Ava I 2. Bam HI 3. Bgl II 4. ECO RI 5. ECO RII 6. Hae III 7. Hha I 8. Hind III 9. Hpa I

Question 34

Organism from which derived: 10. Klebsiella pneumoniae 11. Moraxella bovis 12. Moraxella bovis 13. Providencia stuartii 14. Serratia marcescens 15. Streptomyces albus G 16. Thermophilus aquaticus 17. Xanthamonas malvecearum What are the Enzyme called?

Answer 34

Enzyme: 10. Kpn I 11. Mbo I 12. Pst I 13. Sma I 14. SstI 15. Sal I 16. Taq I 17. Xma I

Question 35

A restriction enzyme with _________________________ recognition sequence **cuts long DNA** more frequently and **produce smaller** _____________than a restriction enzymes with a **six** nucleotide recognition sequence.

Answer 35

* restriction enzyme with a **four** nucleotide * DNA fragments

Question 36

Any given ____________ long recognition site occurs in DNA, on average, at a distance of _____ (4^4) nucleotides. Any given _________________ long sequence occurs, on average, at a distance of _____ (4^6) nucleotides. Any given ______ long sequence occurs, on average, at a distance of _____ (4^8) nucleotides

Answer 36

* four nucleotide long recognition site * 256 * six nucleotide long sequence * 4096 * eight nucleotide long sequence * 65,536

Question 37

_________________ allows scientists to **extract and analyze bits of microbial DNA** from samples, meaning they don't need to find and grow whole cells.

Answer 37

Polymerase Chain Reaction (PCR)

Question 38

**PCR** is an essential element in DNA ________ and in the sequencing of ________ and the entire ____. Basically, it's like a **technique to photocopy pieces of DNA**. In a matter of a few hours, a single DNA sequence can be amplified to ________ of copies. PCR lets scientists work with samples containing even ______ starting amounts of DNA.

Answer 38

fingerprinting genes and entire genomes amplified to millions very small

Question 39

**PRC:** The technique makes use of the **DNA repair** _______. This enzyme, **present in all living things**, fixes breaks or mismatched nucleotides in the ______. These **breaks or mismatches** could cause genes to ____ **if left unfixed.**

Answer 39

enzyme polymerase double- stranded DNA helix malfunction

Question 40

Not all _______ are created equal, however. Many fall apart in _____. **PCR** was developed in ____ following the discovery of an **unusual heat-loving bacterium** called ________ in a hot spring in Yellowstone National Park. This bacterium' s polymerase, **dubbed Taq**, does its job of ______ and __ *nucleotides* even in the **high heat** generated by the successive "____" cycles required during PCR. **Taq made PCR possible**.

Answer 40

polymerases high heat 1985 Thermus aquaticus matching and attaching "photocopying"

Question 41

_____ uses the **intact half of the DNA molecule** as a template and **attaches the right nucleotides**, which ____ constantly in the cell, **to the complementary ____________** at the site of the break. (DNA consists of two strands of nucleotide bases, which are represented as ______. In the laws of DNA base-pairing, __ joins with __ and __ with __.)

Answer 41

Polymerase circulate **nucleotide** A, G, C, and T A joins with T and G with C

Question 42

**Inserting a DNA sample into a Plasmid**: DNA is spliced by ________ pairing and sealed with ______

Answer 42

complementary base DNA ligase

Question 43

Are **small circles of DNA found in bacterial cells**, separate from the bacterial chromosome

Answer 43

Plasmids

Question 44

**cut across the two strands** leaving loose ends to which cDNA can be attached

Answer 44

Restriction enzyme

Question 45

The ____ enters the bacterial cell and **reproduces itself**. When the bacterials cell divides, the ______ are shared cut between the ______ and the it continue to reproduce.

Answer 45

plasmids plasmids two daughter cells

Question 46

Two type of Bioreactors

Answer 46

Batch and Chemostat (CSTR)

Question 47

_____: **changing conditions** - transient (S, X, growth rate), **high initial substrate**, **different phases of growth**. * *more common*

Answer 47

Batch

Question 48

____: **steady-state**, *constant* low concentration of substrate, constant *growth rate* that can be set by setting the *dilution rate* (i.e. the feed flow rate) * more efficient

Answer 48

Chemostat

Question 49

**Choice of continuous vs. batch production**

Answer 49

* Productivity * Flexibility * Control * Genetic stability * Operability * Economics * Regulatory

Question 50

Reactor Choices pt.1 * _________: **rate of product per time per volume**. Chemostat better for growth associated products. Wasted time in batch process. * _________: **ability to make more than one** product with the same reactor. Batch better. * _________: **maintaining the same conditions for all of the product** produced. In theory, chemostat better, steady state. In reality????

Answer 50

Productivity Flexibility Control

Question 51

Reactor Choices pt.2 * _________: **maintaining the organism with the desired characteristics**. Chemostat selects for fast growing mutants that may not have the desired characteristics. * _____: **maintaining a sterile system**. Batch better. * _____: **validating the process**. Initially, many process batch, too expensive to re validate after clinical trials.

Answer 51

Genetic stability Operability Regulatory