Lesson 9: Gene Transfer, Regulation, and Biotechnology

Question 1

Genetic information from two different parents by meiosis

Answer 1

Parasexual

• enhances genetic diversity
• unidirectional from donor to recipient

Question 2

Conjugation was discovered by

Answer 2

Joshua Lederberg and Edward Tatum

Question 3

Carries genes enabling the transfer of DNA from an F positive to an F negative cell
A circular piece of DNA that can exist as an extra chromosomal element or may integrate into the chromosomes of the host cell

Answer 3

Episome

Question 4

Possesses the recombinant DNA in all of its body cells

Answer 4

Transgenic organism

Question 5

Consist of a combination of DNA from more than one gene or more than one species

Answer 5

Recombinant DNA

Question 6

What are the steps in PCR?

Answer 6

• denaturation-DNA template is separated by using heat (95°)
• annealing-DNA primers will bind to each DNA strand based on complementary base pairs(55°)
• extension-DNA polymerase will create a new strand of DNA that is complementary to the template strand forming a primer on each strand

Question 7

Where does heat-stable DNA polymerase come for use in PCR?

Answer 7

Isolated from the thermophilic bacteria (T. aquaticus, and archeae P. furiosus)

*taq polymerase

Question 8

What is needed for PCR?

Answer 8

• DNA template
• taq polymerase
• primers (oligonucliotides)
• nucleotides
• cofactors
• buffers

Question 9

Is real time PCR qualitative or quantitative?

Answer 9

Quantitative

Question 10

• uses two fluorescent dies to collect quantitative data
• same components of PCR
• also requires probe, this allows the researchers to monitor the process of the reaction as it occurs

Answer 10

Real time PCR

Question 11

In real time PCR, the reporter dye is on the _____ and quencher dye is on the ____ end. Called florescence energy transfer.

Answer 11

5’

3’

Question 12

Two newer DNA sequencing methods

Answer 12

• next generation sequencing

- third generation sequencing (• Single molecule, real-time (SMRT) • Nanopore sequencing)

Question 13

DNA that will replicate without integrating into a host chromosome such as plasmid or other types of viruses. Serves as a carrier of the DNA segment to be cloned. Carries from donor to recipient cell

Answer 13

Vector DNA

Question 14

Enzymes that are used to cut genes of interest from a donor cell. Used to digest DNA
Enzymes that cut DNA creating blank cuts or sticky ends , Results in “restriction fragment”
Recognize and cut at palindromes (racecar)

Answer 14

Restriction endonucleases

Question 15

Separates DNA based on size, charge, shape
DNA can then be transferred from gel to nitrocellulose membrane and probes maybe used to localize DNA sequences of interest
After DNA is cut from the donor it can be pasted into recipient DNA (vector) using ligase

Answer 15

Agarose gel electrophoresis

Question 16

Transfer of recDNA to cloning host = “__________”

Answer 16

Transformation

Question 17

Uses electricity to make small holes temporarily in the membrane or wall of the recipient cell

Answer 17

Electroporation

Chemicals can also create temporary holes

Question 18

Use in Vitro fertilization or can entire nucleus be integrated

Answer 18

Micro injections

Question 19

Fatty bubbles that can encapsulate DNA. Brought into the cell by endocytosis or fusion with the plasma membrane

Answer 19

Liposomes

Question 20

This is more common, when you placed the cells in the DNA in the same tube and increased temperature. Transiently allowing the membrane of the cell to be more soluble, then placed on ice to make membrane more rigid to trap the DNA within

Answer 20

Heat shock

Question 21

Encodes beta-galactosidase, cancleave substrate to turn colony blue

Answer 21

lacZ

Question 22

Steps for the production of recombinant DNA

Answer 22

• Purify chromosome DNA
• purify the plasmid (vector)
• insert the gene of interest into the vector
• transformation
• Selection of transform cells

Question 23

Nitrocellulose membrane, a piece of nucleic acid is used as a probe to detect the DNA and bind

Answer 23

Gel electrophoresis

A way to analyze DNA

Question 24

Separated DNA fragments are transferred from gel to nitrocellulose membrane.
Probes used to localize DNA sequence of interest

Answer 24

Analysis of DNA by Southern blot

Question 25

Applications of southern blotting/DNA fingerprinting

Answer 25

Analyze evolutionary links, study of endangered species, tell he was related to whom, identify specific genes, diagnosis of infectious disease, detect pathogen’s in food samples, detect genetically modified foods or organisms, forensic analysis

Question 26

Used to identify specific RNA molecule, it can determine if the specific gene is transcribed. Example is a nerve cell versus a muscle cell. Also determine if pre-mRNA is alternatively spliced. Example fetal cells versus adult cells

Answer 26

Northern blotting

Question 27

• used to identify specific proteins

- Specific proteins are detected by antibodies. If a protein made at a particular stage of development

Answer 27

Western blotting

Question 28

What is a DNA hybridization test?

Answer 28

Probes made of nucleic acid‘s bind complementary regions
-reporter Molecules (•Fluorescent dyes, • enzyme linked)
Used to diagnose cause of disease, identify pathogens and genetic disorders

Question 29

It has that increase his temperature to denature into single strand and then decreases temperature to produce two strands to anneal, different strands with compatible base pairs also aneal only Homologous DNA will attach

Answer 29

DNA hybridization tests (nitrocellulose)

Question 30

Test used to detect the physical location on a gene

Answer 30

In-situ hybridization test (FISH) (Florence used)

Question 31

• Probes apply to intact cells
• observed microscopically for the presence and location of specific genetic marker sequences
• effective way to locate genes on chromosomes “Genetic Mapping”
• ex: Test for leukemia

Answer 31

Analysis of DNA by fluorescent in situ hybridization (FISH)

Question 32

Molecules of immobilized single-stranded DNA fluorescently labeled molecules that are washed over the micro array
-Used to screen for inherited diseases, diagnose infection, monitor gene expression

Answer 32

Genetic screening by using DNA microarrays

Question 33

Can somatic cell gene therapy be inherited by the offspring?

Answer 33

No, it cannot help the offspring
This helps with symptoms but does not cure disease
Introduces wild type allele into mutant cells via a viral vector

Question 34

This type of gene therapy could cure disease and prevent transmission
Ethical problems that could be induce from the therapy most problems are trace to the vectors used

Answer 34

Germ line gene therapy

This is heritable

Question 35

These often have transient expression because the gene is not integrated into the chromosome therefore the gene is only expressed as long as the viral infection persist

Answer 35

Adenoviral vectors

Question 36

This type of gene therapy randomly integrate causing mutations in the gene insertion cannot be controlled

Answer 36

Retrovirus gene therapy