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Flashcards in Chapter 8 genomics Deck (35):
1

In biology, this is the process of producing populations of genetically-identical individuals

Cloning

2

The process of creating copies of DNA fragments, and sometimes, the protein products of the DNA

Cloning (for us in genetics)

3

What is essential to study genomes?

Cloning

4

What do tumors need to grow and spread

blood supply

5

This is a cloned antibody tht inhibits vascular endothelial growth factor (VEGF), which promotes new blood vessels (angiogenesis)

Avastin

6

This is used to treat colon, lung, and breast (formerly) cancers

Avastin

7

_______is a positive key to making massive quantities of a drug

Cloning

8

Basic Cloning Tools

1. DNA from organism of interest
2. Restriction enzyme (endonucleases
-cut DNA at specific recognition
3. DNA ligase
-joins recombinant DNA molecules permanently
4. Cloning vectors
-Vehicle for replication of foreign DNA
5.Host microorganism (often bacteria)
-Factory for propagation of recombinant DNA in vector

9

Occur naturally in bacteria-purpose is to cut viral DNA that infects genome, thus restricting access of the virus

RE's (restriction enzymes)

10

about how many RE'S are known

400

11

recognize specific DNA sequences in the genome called _____ ____ which are often palindromes-mirror images of same sequence

restriction sites

12

cleave phosphodiester bonds, leaving 5' phosphate and 3' hydroxyl group in the extracted fragments

RE'S

13

_______ DNA protects bacterial genome from RE'S

Methylated DNA

14

RE probability is

(1/4)^n where n=number of bases

15

RE cuts both strands of DNA between same two base pairs

Blunt Ends

16

RE'S make staggered cuts in DNA

Sticky (overhanging) ends

17

Why are sticky ends useful

1. fragments of DNA cut by same RE with same sticky ends bond with each other easily.
2. ligase cements these bonds
3. new DNA is molecule is recombinant

18

How do we make lots of copies of recombinant DNA?

plasmid DNA vectors

19

plasmid vectors are engineered to have

1. an ori sequence needed for independent replication in bacteria.
2. a selectable marker to identify bacteria with the plasmid (e.g. antibiotic resistance).
3. At least one unique restriction site for insertion of recombinant DNA.

20

RE'S have restriction sites of different lengths which are .......

4, 6, and 8, maybe more 6 has the most

21

PCR was not a discovery but rather an invention

Polymerase Chain Reaction

22

BORN in 1944 the inventor of PCR was awarded 1993 Nobel Prize in Chemistry

Kary Mullis

23

Founded a company to sell jewelry with DNA of famous dead people like Elvis and Marilyn Monroe

Kary Mullis

24

talked to a raccoon and believes in extraterrestrial visitors to earth including a glowing green raccoon

Karry Mullis

25

Can be used to make many copies of any DNA that is supplied as a template

PCR

26

Starting with one original copy an almost infinite number of copies can be made using

PCR

27

"Amplified" fragments of DNA can be sequenced, cloned, probed, or sized using electrophoresis with _____

PCR

28

Why is PCR very important #1

1. cloning for DNA sequencing
2. DNA-based phylogeny

29

Why is PCR very important #2

1. Diagnosis of genetic disease
2. diagnosis of infectious disease such as swine flu.
3. Genetic fingerprinting (forensics & paternity testing

30

what is MCS

Multiple cloning site

31

MCS is located in a gene called

beta-galatosidase

32

How PCR works

1. PCR is an artificial way of doing DNA replication.
2. Instead of replicating all the DNA present, only a small segment is replicated, but this small segment is replicated many times.
3. As in replication, PCR involves:
-Unwinding DNA (melting in PCR)
-Priming
-Polymerization

33

Components of a PCR reaction

1. Buffer (containing Mg++)
2. Template DNA
3. 2 Primers that flank the fragment of DNA to be amplified.
4.dNTP's
5. Taq DNA Polymerase

34

What happens if an error occurs early in the PCR process?

the mistake can be magnified millions of times, giving a mistake in the sequence

35

Draws of PCR are

1.regular Taq polymerase has NO proofreading ability, but some different kinds of DNA polymerase for PCR can minimize error rates.
2. If a mismatch error occurs early in the PCR process, the mistake can magnified million of times, giving a mistake in the sequence.