Chapter 20

Question 1

biotechnology

Answer 1

the manipulation of organisms or their components to make useful products

Question 2

genetic engineering

Answer 2

the direct manipulation of genes for practical purposes

Question 3

nucleic acid hybridization

Answer 3

key to many of the DNA techniques/tools (base pairing to complement = basis)

Question 4

completed extinct genomes

Answer 4

wooly mammoth and neanderthals

Question 5

DNA sequencing

Answer 5

a complete nucleotide sequence of a DNA molecule

1st based on dideoxy (chain termination sequencing)

Question 6

dideoxy

Answer 6

the fragment of DNA is denatured to single strand and incubated with DNA ingredients

modified dNTPs added (fluorescent) -> added beginning 3’ until adding themselves

Question 7

“Next-generation sequencing”

Answer 7

thousands of fragments sequenced in parallel
one nucleotide at a time
electronic monitors

Question 8

How many nucleotides do women have?

Answer 8

approx 3 bill (completed using dideoxy)

Question 9

What was a major thrust for the Human Genome Projeccr?

Answer 9

faster and cheaper sequencing

Question 10

whole-genome shotgun approach

Answer 10

cloning and sequencing of fragments of randomly cut DNA followed by assembly into a single continuous sequence

Question 11

map-based sequencing

Answer 11

recombination studies - overlapping DNA

Question 12

DNA cloning

Answer 12

well-defined DNA segments in multiple identical copies

Question 13

two types of cloning

Answer 13

gene cloning (the production of multiple copies of a single gene)

organismal cloning

Question 14

Why clone just one gene?

Answer 14

agricultural improvements (pest-resistance)

environment (modify bacteria to clean)

medicine (like insulin)

Question 15

plasmids

Answer 15

small circular DNA molecules that replicate separately from the bacterial chromosome

Question 16

recombinant DNA

Answer 16

a molecule with DNA from two different sources

Question 17

cloning vectors

Answer 17

area copied to create identical genes

Question 18

Why are bacterial plasmids widely used as cloning vectors?

Answer 18

They are readily obtained, easily manipulated, easily introduced into bacterial cells, and once in the bacteria they multiply rapidly

Question 19

Generally plasmid vector contains:

Answer 19

origin of replication

genes for antibiotic resistance

multiple cloning sites

Question 20

restriction enzymes

Answer 20

cut DNA molecules at specific DNA sequences called restriction sites (making restriction fragments)

Question 21

DNA ligase

Answer 21

an enzyme that seals the bonds between restriction fragments

Question 22

polymerase chain reaction (PCR)

Answer 22

produce many copies of a specific target segment of DNA

Question 23

transformation

Answer 23

inserting a new piece of DNA into bacteria

Question 24

How will we know the bacteria took up the plasmid?

Answer 24

the antibiotic resistance gene (amp)

Question 25

How do we know which bacteria are carrying a plasmid with our gene of interest?

Answer 25

blue/white colony screening

Question 26

expression vector

Answer 26

a cloning vector that contains a highly active bacterial promoter

Question 27

cDNA (complementary DNA)

Answer 27

researchers avoid the intron problem with cDNA complementary to the mRNA (template for PCR amplification)

Question 28

How do you avoid eukaryote-bacterial incompatibility?

Answer 28

yeast, insect, or mammalian cells

Question 29

electroporation

Answer 29

applying a brief electrical pulse to create temporary holes in plasma membranes

Question 30

In situ hybridization

Answer 30

uses fluorescent dyes attached to probes to identify the location of specific mRNAs in place in the intact organism

Question 31

Reverse transcriptase-polymerase chain reaction (RT-PCR)

Answer 31

useful for comparing amounts of specific mRNAs in several samples at the same time

Question 32

organismal cloning

Answer 32

produces one or more organisms genetically identical to the “parent” that donated the single cell

Question 33

nuclear transplantation

Answer 33

the nucleus of an unfertilized egg cell or zygote is replaced with the nucleus of a differentiated cell

Question 34

Are clones always exact?

Answer 34

No

Question 35

stem cells

Answer 35

relatively unspecialized cells: produce indefinitely and potentially differentiate into one or more specialized cell types

Question 36

embryonic stem cells

Answer 36

produce indefinitely

Question 37

adult stem cells

Answer 37

can generate multiple cell types and are used in the body to produce nonreproducing cells