Ch 9 Flashcards
A) What are restriction enzymes? What do they do?[Figure 9.1, Table 9.2, Restriction endonuclease video*]
cut dna into fragments . each recognize 4-6 nucleotide sequence.
B) What are restriction fragments? How do they relate to restriction enzymes?
q
C) What are recombinant DNA molecules? [Figure 9.1]
molecules made by joining dna from 2 different sources.
D) Define the term anneal.
forming base pairs
E) What does DNA ligase do? [Figure 7.7 step 5 (pg167)]
enyme used to form the covalent bonds between adjacent nucleotides
F) What is gel electrophoresis? How is it done? What sort of things do scientist use it for?[Figure9.2]
dna fragments are funneled through via electrical current( gel is a conductor, dna is slightly neg. and they are pulled to a positively charged electrode). The different length fragments are caught through the funnel. This provides a “finger print of the dna”
A) Review table 9.3
??????? pg 218
B) What is DNA cloning? [Figure 9.3, For further reference pgs. 166, 201 and 208]
restriction enzymes cut portion of dna which is then transferred into another organism. The dna transfered must be able to replicate in order for future generations to be produced. Most do not contain the orgin of replication and therefore cannot replicate, but that is why you add it to a vector, or plasmid.
C) What sort of proteins are produced by genetically engineered microorganisms? How are they used?
insulin. These are protiens that used to be extracted from live animals , but are now produced by gmo micro organisms. This helps create vaccines, renin in cheese production
D) What is “shotgun cloning” ?
e coli has fewer genes than most and so it is key in helping find what a particulair gene produces by introducing the gene into it.
E) What is a reporter gene? How is it involved in gene fusion? What is gene fusion?[Figure 9.5]
a gene that’s product is easy to see. this is taken and is joined to the gene to make it visible and obvious when that gene is incorperated into another microorg.
F) What does it mean if someone says an animal is transgenic? What does the Ti plasmid have to do with it? Why can transgenic plants be a good thing?
It harbors a cloned gene.
Ti plasmid is used as a vector to transfer gene information.
can be used to transfer genes that act as natural insecticides rather than toxic chemicals being used. Also can improve their nutritional value. Also vaccinations via food
A) What is a DNA library? How is it created? [Figure 9.6]
a collection of clones that together make an entire genome. take restrictive enzyme, then place each separate gene into diff ecoli. then you can see which one creates the protein you want.
B) When you are going to clone DNA what is the first step and how is it accomplished?
obtain dna you wan to be cloned
adding getergent to lyse the cell
in eukaryotic cells requires mrna then used that to make the dna because original dna has introns
C) What is cDNA? [Figure 9.7 and cDNA video *]
copy of dna made from mrna strands without introns
D) What is a vector? [Figure 9.8 and Construction of a plasmid vector video *]
modified plasmid or bacteriophage
has orgin of replication and function as carriers of cloned dna.
E) What is a selectable marker? Why is it important?
gene that allows vector to growin otherwise inhibitory or lethal conditions. This allows researchers to know which cells took up the dna!
F) What is the lacZ’ gene? What is it used for? [Figure 9.9]
Lac z is a gene found in a second genetic marker puc 18. It’s product cleaves to a colorless chamical xgal to form a blue compound.
G) What is DNA-mediated transformation?
when they use electroporation to get a cell to take up dna
H) What is electroporation?
this is when they shock the cell causing it to take in the dna
A) Why do some people fear GM foods? What precautions are being taken?
There are people who are afraid that new genes can cause unexpected allergic reactions. This is why they are monitored by the FDA.
A) What is DNA sequencing?
Process of determmining nucleotide sequence of a dna molecule
B) What is the Human Genome Project? How has there research benefited science?
This project was to find the human genome sequence which led to then discovering prokaryotes and eukaryotes sequences . This has created a lot of data and helps us to know relations/ connections between organisms
C) What is the Human Microbiome Project? What are it’s goals?
using genomes to discover variations of different variations in normal microbiota in humans. and comparing them in health and disease.
D) What is the dideoxy chain termination method? What is it used for? [Figure 9.12 (Know the steps), Figure 9.13]
This is when it terminates in invitro dna synthesis reaction has 5 parts
Template dna
Polymerase: synthesizes dna
Primer: Short dns sequence complimentary to template dna adheres to it providing an idea of where researcher wants synthesis to start.
Deoxy nucloetides used in dna synthesis
dideoxynucleotides: almost same as deoxynucleotieds, but lack 3 oh group
A) What is template DNA? [Reference pg. 163]
template dna is original piece of dna that is used to make copies etc.
B) What is DNA polymerase? [Reference pg. 166]
This is synthesizes dna
C) What is a primer? [Figure 9.10 and reference pg. 166]
a short complimentary strand that adheres to the template dna
D) What are chain terminators? What do they do? [Figure 9.11]
deoxynucloetides are chain terminators. They prevent addition of more nucleotides to dna sequence
A) What is PCR? [Figure 9.14 and Polymerase chain reaction video *]
allows million copies of certain section(target dna) of dna to be replicated. and is used to detect diseases
A) How does double-stranded DNA relate to PCR?
this is the template used
B) How does Taq polymerase relate to PCR?
heat stable polymerase
C) How does primers relate to PCR?
allows researchers to choose where syntheis starts
D) How does deoxynucleotides relate to PCR?
4 of them which are used in synthesis
B) What are the three steps in the PCR repeating cycle? [Figure 9.15]
- smple is heated to boiling point denaturing protien
- then it is quicly cooled causing the primers to aneal to the denatured target dna
- raised to optimal tac polymerase temp and allow synthesis to occur.
C) What is the PCR product? How is it formed? [Figure 9.16 and figure 9.17]
fragment containing target dna is the product. Fromed via three cycles of pcr. the length of the target dna is made via replication of medium dna (2nd cycle)
pg 230
A) What is a DNA probe? [Figure 9.18]
is a single strand of dna with a sequence of desired target dna( this is connected to a radioactive isotope or florescent marker.
B) What is hybridization?
when probe anneals to the dna strand
C) What is colony blotting? What is it used for? How is it done? [Figure 9.19]
pg 233
nylon is placed over dish and then it takes up the colonies. THen it is dipped in alkaline solution lysing the cells, and then a probe is added . This helps you locate which colonies have the desired dna
D) What is fluorescence in situ hybridization? How is it done? What is it used for? [Figure 9.20]
using florescents to mark cells containing desired dna. The probes are usually targeted for specific rrna in prokaryotes. this is because there are typically more of them
E) What is a DNA microarray and what is it used for? [Figure 9.21 and Microarray video*]
study gene spression of genomes which have been sequenced. ????? pg 233
