Ch. 9 Biotech

Question 1

What is biotechnology

Answer 1

of microorganism to make a product

Question 2

What is recombinant DNA?

Answer 2

r(DNA) insertion or modification of genes to produce wanted proteins through a vector for example

Question 3

Vector

Answer 3

A DNA molecule that transports foreign DNA into a cell. The DNA molecule HAS to be self-replicating, such as a plasmid or viral genome.

Question 4

Clone

Answer 4

clones of the cell that has the vector for the purposes of DNA

Question 5

Examples of biotechnology?

Answer 5

They have done this for making bacteria when for insect eat plants they die, and for medications , or we can makee a genen that contains a degrative enzyme that canbe sued to clean up toxic waste, we can use it to make e coli make hGH humaan growth hormone

Question 6

site-directed mutagenesis

Answer 6

targeted and specific change to a gene

Question 7

Restriction enzymes

Answer 7

tools used for manipulation (we’ve exploited them to molecular scissors)
They cut specific sequences of DNA. They hydrolyze phage DNA in order to protect themselves, but they cannot kill themselves because they methylate some of their cytosines in their DNA

Question 8

What restriction enzyme did we make from E.coli?

Answer 8

EcoR1

Question 9

What is a blunt end?

Answer 9

When a restriction enzymes cuts at two places on the same strand

Question 10

What is a sticky end

Answer 10

The end of the cut DNA by a restriction enzyme with some parts sticking out, these are better for rDNA because they introduce a plasmid that contains the complementary base paring

Question 11

What is PCR?

Answer 11

Polymerase chain reaction (PCR) Used to make copies of DNA enzymatically. Clone DNA, amplify DNA number for detection. Diagnose genetic Disease and detect pathogens . Use heat to separate strands, then add the pieces needed to remake them primers, nucleotides, and DNA polymerase and make more, rinse and repeat

Question 12

Why are shuttle Vectors important

Answer 12

They can survive in several different species, can be use to move cloned DNA to say bacteria to fungi etc.

Question 13

Why can viral DNA be a useful vector

Answer 13

they can accept larger DNA than plasmids

Question 14

What is the method used to shock DNA into a cell

Answer 14

Electroporation, used to insert DNA into a cell

Question 15

What is the method called to introduce DNA into a cell via vectors

Answer 15

Transformation , they take DNA from the environment

Question 16

Method where you enzymatically remove cell wall

Answer 16

Protoplast fusion, no cell wall means easier access. Then it can melt into another cell. this is useful for plants and algal cells

Question 17

What is a gene gun for

Answer 17

Shoot DNA through the cellulose walls of plant cells, another method of DNA introduction

Question 18

Method where you inject DNA into an animal cell

Answer 18

microinjection, because the needle is tiny

Question 19

What is a genomic library

Answer 19

Collection f bacteria that each have different pieces of the genome of say an animals. Either in a phage (viral) library or plasmid library.

Question 20

Why is eukaryotic DNA hard to clone genes from?

Answer 20

Has protein coding parts exons , and non protein coding parts introns, The introns make it big and bulky, and normally are taking out when converted to RNA, bacteria can’t always take them out themseslves. so an artificial gene is made that only contains exons through reveres transcriptase.

Question 21

what is cDNA

Answer 21

Complimentary DNA that comes from mRNA, and has the introns and exons

Question 22

What is synthetic DNA

Answer 22

made via DNA synthetic machines, much like typing an essay, but only up to 200 sequences and they need to be stitched together, and you need to know the instructions

Question 23

What is a DNA library

Answer 23

collection of DNA seq that have been put into plasmids , So one colony would have a certain segment of DNA,

Question 24

Why do we use E.coli a lot and what are disadvantages?

Answer 24

s easily grown and genomics are well known
But endotoxin must be removed from their outer cell wall. it causes fever and shock, and the cells need to be broken up in order to harvest proteins from there. it does not secrete them.

Question 25

What other microbe is used in biotechnology and why?

Answer 25

Bacillus subtilis because it secretes protein. and are gram positive. Saccharomyces cerevisiae (yeast), its genome is 4 times bigger but it a well understood eukaryotic cell. they may be more successful in creating the product as well as continuously secreting them

Question 26

What are mammalian cells used for

Answer 26

They can be used for making protein products for medical use because they secrete them and theres lower risk of toxins and allergies. For example CSF that stimulate the growth of cells that protect from infection. Normally we make little on our own.

Question 27

What are plant cells used for

Answer 27

plants cells are altered by rDNA to make products like alkaloids (pain killers) rubber, sunscreen,. In order to modify them you also need bacteria sometimes.

Question 28

what is a subunit vaccine

Answer 28

made through rDNA, a subunit vaccine only contains the protein part of a pathogen

Question 29

Dna vaccines

Answer 29

made through rDNA, plasmids carry a viral gene under the control of a poromote active in human cells. its hard to get the plasmid DNA into human cells. Also we can use it for makeing human enzymes and hormones’

Question 30

Gene therapy

Answer 30

rDNA allows for replacing defective genes or missing genes

Question 31

Gene silencing

Answer 31

suppresses certain genes siRNA (small interefineg) bind to mRNA which is then destroyed by RISC thus silencing the expression of gene

Question 32

Shotgun sequencing

Answer 32

A genome project where small pieces of genome in a living cell are sequenced and then assembled using a computer . Can be used to study microgorganisms that have not been cultured.

Question 33

Human genome project

Answer 33

a project where they sequenced all og human DNA and hope it will provide info on diagnostics and treatments. Through blocking a gene we can determine its function (reverse genetics ). foward genetics is starting wwith pehnotype and looking for the genotype

Question 34

What is it called determining the proteins made by a cell

Answer 34

proteomics

Question 35

What is southern blotting

Answer 35

testing to see if a certain gene is present. can be used for forensics. band on the filter highlights the gene

Question 36

Real-time PCR

Answer 36

newly made DNA is tagged with fluorescent dye. levels of florescence can be measured after every PCR cycle.

Question 37

reverse transcription RT-PCR

Answer 37

is using reverse transcriptase by getting DNA from RNA or mRNA

Question 38

How is it used in forensics

Answer 38

evidence collection, rape conviction ,tracing HIV , anthrax in mail etc. DNA fingerprinting can be used to identify bacterial or viral pathogenic

Question 39

Why is agrobacterium important

Answer 39

it has a TI plasmid, that occurs naturally within it. as a result with rDNA we can make it pest resistant, herbicide resistant, suppressed gene that prevents enzymes from breaking down pectin, and thus have a longer shelf life or more nutritious

Question 40

what are some ethical issues to look out for

Answer 40

accidental release, modified crops have to be safe for ppl and environment. Who has the righ tto have access to a persons genetic info?