recombinant DNA

Question 1

how are some disease causes

Answer 1

by individuals unable to make certain protiens

Question 2

how are protiens created

Answer 2

by the lengths of dna

Question 3

how were mising protiens fixed previously

Answer 3

the protien was taken from an individual and injected into the host

Question 4

what issues did this cause

Answer 4

rejection

Question 5

what has technology allowed

Answer 5

us to isolate the gene , manipulate them and insert into the organism

Question 6

what is recombinant dna

Answer 6

when the dna from two species are combined forming gmo or transgenic

Question 7

why can we produce recombinant dna

Answer 7

as the genetic code is universal so transcription and translation can occur in all cells

Question 8

what are the summary stages

Answer 8

isolation of dna 
insetion of isolated dna into vector
transformation into suitable host
identification of sucessful host
growth and cloning of succesful host

Question 9

how can we isolate a gene

Answer 9

by converting mrna to dna using reverse transcriptase
using restriction endonucleases to cut fragmented gene
make gene using the gene machine

Question 10

what is recognised first

Answer 10

the cell which produces the required protien

Question 11

what happens to the cell

Answer 11

it is centrifuged to release MRNA

Question 12

what happens to the mrna

Answer 12

with reverse transcriptase it forms a complimentry dna strand forming a hetro duplex

Question 13

what happens to this hetroduplex

Answer 13

it is hydrolysed releasing the mrna strand

Question 14

what happens to the resulting dna strand

Answer 14

the complementry dna numcleotides join using dna polymerase

Question 15

why do bacteria have restriction endonuclese

Answer 15

to protect them from invading virus they are able to cut the dna at a certain section

Question 16

what do the enzymes have

Answer 16

active sites that cut as specific recognition sites

Question 17

how are the recognition sits usiually cut

Answer 17

in a palindrome way with sticky ends at opposite base pairs

Question 18

what are the two types of ends

Answer 18

blunt or staggered sticky ends

Question 19

what do these sticky ended fragments contain

Answer 19

the dna for the desired gene

Question 20

what is determined in the gene machine

Answer 20

the sequences of bases for the desired protien aswell as its amino acid sequence

Question 21

what triplets are worked out

Answer 21

the complementry dna amino acid triplets

Question 22

what is done to this sequence

Answer 22

it is fed into a computer

Question 23

what does the computer design

Answer 23

small sequence of overlapping single nucleotide strands

Question 24

how are the strands assembled

Answer 24

one base at a time into the correct sequence to make the desired gene

Question 25

what does the gene contain/ not contain

Answer 25

it only contains extrons

Question 26

how is the gene replicated

Answer 26

using the polymerase chain reaction

Question 27

how are the right genes checked

Answer 27

they are scrrened under standard sequencing techniques with errors being rejected

Question 28

in reverse transcriotase methods the complimentry dna contain only what

Answer 28

exons so is shirter than other dna

Question 29

what is the strength of in vivo cloning in terms of contamination

Answer 29

less likely to get contaminates as only bases with complimentary base pairing can anneal which the bases are cut using restriction enzymes

Question 30

what also does it produce

Answer 30

produces an active protein rather than just the gene as the bacteria will transalate the gene in its genome

Question 31

what is the disadvantage of in vivo

Answer 31

it is time consuming as the gene has to be identified inserted and transformed and then cloes

Question 32

what else is a weakness in terms of succession

Answer 32

transformation has a very low success rates and recombinace is very low success

Question 33

what is needed

Answer 33

pure large amount of dna