DNA recombination and technology

Question 1

Restriction enzymes

Answer 1

endonucleases produced by bacteria
recognize specific 4-to 8-bp short
palindromic sequences(restriction sites)cleave both DNA strands at this site.
It recognizes and cleave foregin DNA
protects DNA from the DNA of foreign organisms by digestion

Question 2

PCR-RFLP

Answer 2

Restriction Fragment Length Polymorphism

inherited difference in homologous DNA detected with fragments of different length

Question 3

DNA library

Answer 3

Fragments of an entire genom cloned into vectors

Question 4

cDNA library

Answer 4

Fragments prepared from total DNA of cell or tissue and comprises copies of mRNA

Question 5

Polymerase chain reaction (PCR)

Answer 5

Denaturation, annealing, elongation

Question 6

Tools for PCR

Answer 6

dNTP, primer, Dna polymerase(thermostable), template

Question 7

Blotting teqniques

Answer 7

RNA-nothern
DNA-soutern
protein-western

Question 8

STR

Answer 8

short tandem repeat

Question 9

VNTR

Answer 9

variable number of tandem repeats

Question 10

SNP

Answer 10

single nucleotide polymorphism

Variation in a single nucleotide that occurs at a specific position in a genome

Question 11

Viral vectors

Answer 11

Retroviral vectors
Adenosine deaminase deficiency (ADA)
Severe combined immunodeficiency (SCID)
Adenoviral vectors
Adeno-associated viruses (AAV)

Question 12

Non-viral vectors

Answer 12

Liposome-based

Modified transposons

Question 13

Two Enzymes involved in cloning vectors

Answer 13

• Restriction Enzymes

- T4 DNA Ligase

Question 14

LTR

Answer 14

Enhancer, promoter, transcription initiation (capping), transcription terminator and polyadenylation signal. Expression directed by the viral LTR signals is carried out entirely by host cell enzymes (RNA pol II, poly A synthetase, guanyl transferase).

Question 15

Retionblastoma

Answer 15

tumosupressor

Question 16

ASA-PCR

Answer 16

4 primers, added to 3´-end closest to mutation

Question 17

DNA protected from digestion by

Answer 17

metylation catalyzed by DNA methylase

Question 18

DNA ligase

Answer 18

catalyzes formation of phosphodiester bond with ATP as cofactor

Question 19

Plasmid

Answer 19

circular DNA
own replication
5000–>400 000 bp
resistant to antibiotics
has ori
covalentely bonded to bacterial chromosom
Naked dna and do not code genes necessary to encase the genetic material for transfer to a new host

Question 20

Fingerprinting

Answer 20

based on swquence polymorphism

focuses on differences in lengths of short tandem repeats (STR)

Question 21

FISH

Answer 21

fluorescence in situ hybridization, used to map location of specific DNA sequences witin fixed nuvlei

Question 22

CRISPER

Answer 22

DNA gene editing regulatory system. Providing resistance to external genes from bacteriophage. It combines with The Cas and nuclease to target and cleave the DNA of invading phage.

Gene deletions
gene editing
modulation of gene transcription

Question 23

Cosmid

Answer 23

A plasmid into wich DNA from bacteriophage lambda have been inserted, permits DNA packing in vitro.

Question 24

vector

Answer 24

A plasmid or bachteriophage where foreign DNA can be inserted for purpose of cloning

Question 25

RNA-inducable interferon

Answer 25

Provide same protection against RNA lige restriction enzymes.

Question 26

restriction enzymes

Answer 26

Only present in cells that have enzyme that methylates host DNA–>making it unsuitable substrate for digestion
Site-specific DNA methylases
This enzyme and other exist in pairs

Question 27

Sticky ends

Answer 27

usefull for constructing hybrid or chimeric DNA

MAy reconnect with thelselves, no net gain of DNA

Question 28

Blunt ends

Answer 28

can be ligated directley,

ligation is not directional

Question 29

Homopolymer tailing

Answer 29

A process how Dna anneal with vector. Poly G added to 3´-end of vector and poly d added to 3´endof foreign dna using enzyme terminal transferase.

Question 30

Direct blunt end ligation

Answer 30

bacteriophage T4 DNA ligase and can join any pairs of ends

Question 31

Cohesive ends

Answer 31

can be added through PCR amplification

Question 32

Clone

Answer 32

large population of identical molecules arising from common ancestor
Chimeric or hybrid DNA can be constructed in cloning vectors