Lecture 10 - Genetic Engineering and Recombinant DNA

Question 1

No product or application is directly derived from it

Answer 1

basic science

Question 2

Applications from basic research science

Answer 2

applied science

Question 3

• Clip DNA crosswise at selected positions
• Recognize foreign DNA
• Capable of breaking the phosphodiester bonds between adjacent nucleotides on both strands of DNA
• Protect bacteria and archaea from bacteriophage or plasmids

Answer 3

restriction endonuclease

Question 4

• Each endonuclease recognizes a sequence of 4 – 10 base pairs
• Recognize and clip at palindromes

Answer 4

restriction endonucleases

Question 5

Staggered symmetrical cuts that leave short tails

Answer 5

sticky ends

Question 6

Pieces of DNA produced by restriction endonucleases

Answer 6

restriction fragments

Question 7

• Differences in the cutting pattern of specific restriction endonucleases give rise to restriction patterns of different lengths
• Allows direct comparison of DNA of two different organisms at a specific site

Answer 7

Restriction fragment length polymorphisms (RFLPs)

Question 8

• Necessary to seal sticky ends together by rejoining the phosphate-sugar bonds cut by endonucleases
• Main application is final splicing of genes into plasmids and chromosomes

Answer 8

ligase

Question 9

Replicates HIV and other retroviruses

Able to convert RNA into DNA

Answer 9

reverse transcriptase

Question 10

• Made from messenger, transfer, ribosomal, and other forms of RNA
• Useful in synthesizing eukaryotic genes from mRNA transcripts
• Gene is free of introns

Answer 10

complementary DNA

Question 11

Positions of DNA fragments are determined by staining the DNA fragments in the gel

Answer 11

gel electrophoresis

Question 12

Useful in characterizing DNA fragments

Allow for comparison of genetic similarities among samples in a genetic fingerprint

Answer 12

electrophoresis

Question 13

Two different nucleic acids can ____ by uniting at their complementary regions.

Answer 13

hybridize

Question 14

ssDNA can hybridize with

Answer 14

ssDNA or RNA

Question 15

RNA can hybridize with

Answer 15

other RNA

Question 16

A short stretch of DNA of a known sequence that will base-pair with a stretch of DNA with a complementary stretch of DNA if it exists in a sample

Answer 16

gene probes

Question 17

carry reporter molecules such as fluorescent dyes so that areas of hybridization can be visualized

Answer 17

gene probes

Question 18

detected when non pigmented substrates become color molecules by the action of the enzyme

Answer 18

enzyme linked probes

Question 19

Diagnosing the cause of an infection from a patient’s specimen
Identifying a culture of an unknown bacterium or virus

Answer 19

gene probes use

Question 20

DNA is isolated, denatured, placed on an absorbent filter, and combined with a microbe-specific probe
Blot is then developed and observed for areas of hybridization

Answer 20

hybridization

Question 21

Probes are applied to intact cells

Observed microscopically for the presence and location of specific genetic marker sequences on genes

Answer 21

FISH

Question 22

A very effective way to identify genes on chromosomes
Also effective in identifying bacteria living in natural habitats without culturing them
Used to detect RNA in cells and tissues

Answer 22

FISH

Question 23

Rapidly increases the amount of DNA in a sample without the need for making cultures or carrying out complex purification techniques.

Answer 23

PCR

Question 24

Can detect cancer from a single cell

Can diagnose an infection from a single gene copy

Answer 24

PCR

Question 25

uses the same events of DNA replication

Answer 25

PCR

Question 26

synthetic oligonucleotides of a known sequence of 15 – 30 bases that indicate where amplification should begin

Answer 26

primers of PCR

Question 27

High temperatures necessitate use of DNA polymerases isolated from thermophilic bacteria

Answer 27

DNA polymerase of PCR

Question 28

3 steps of PCR

Answer 28

denature priming extension

Question 29

two good types of cloning vectors

Answer 29

plasmids | bacteriophages

Question 30

- small, characterized, easy to manipulate - can be transferred via transformation - carry genetic markers for resistance to antibiotics

Answer 30

plasmid vector

Question 31

natural ability to inject DNA into bacterial hosts through transduction

Answer 31

bacteriophageRecombinant organisms produced through the introduction of foreign genes Also known as genetically modified organisms (GMOs)

Question 32

Recombinant organisms produced through the introduction of foreign genes (GMOs)

Answer 32

transgenic organisms

Question 33

creation of organisms from scratch

Answer 33

synthetic biology

Question 34

a result of synthetic biology

Answer 34

XNA

Question 35

- Contain different sugars than DNA and RNA, or in some cases, no sugars at all - Enzymes were able to copy resulting in the ability to store information and pass it on to descendant cells. - These findings imply that life on other planets does/did not require DNA or RNA to exist

Answer 35

XNA

Question 36

give an exact order of bases in a plasmid, chromosome, or genome

Answer 36

sequence maps

Question 37

The whole genome of an organism is broken down into smaller, manageable fragments Fragments are separated through gel electrophoresis (inserted into plasmid and cloned into an E coli cell)

Answer 37

whole genome shotgun sequencing WGSS

Question 38

Plasmids are purified and DNA fragments are sequenced by automated sequencers A computer program takes all the sequence data and is able to find where the sequence overlaps

Answer 38

WGSS

Question 39

a larger, contiguous set of nucleotide sequences produced by the automated process

Answer 39

contigs

Question 40

Sophisticated computers and systems developed to sequence genomes in a shorter period of time with less human effort

Answer 40

high-throughput sequencing

Question 41

Manage the data produced through high-throughput screening Analyze and classify genes Determine protein sequences Ultimately determine the function of the genes

Answer 41

genomics and bioinformatics

Question 42

Determining the functional information of a gene sequence

Answer 42

annotating the genome

Question 43

Based on the principle that the way nucleotides are combined is different for each organism

Answer 43

dna profiling

Question 44

only a single nucleotide is altered

Answer 44

SNP

Question 45

Allows biologists to view the expression of genes in any given cell Track the expression of thousands of genes at once

Answer 45

microarray analysis

Question 46

study of the proteins a cell produces at a defined point in time

Answer 46

proteomics