Producing DNA fragment

0.0(0)
studied byStudied by 0 people
learnLearn
examPractice Test
spaced repetitionSpaced Repetition
heart puzzleMatch
flashcardsFlashcards
Card Sorting

1/17

encourage image

There's no tags or description

Looks like no tags are added yet.

Study Analytics
Name
Mastery
Learn
Test
Matching
Spaced

No study sessions yet.

18 Terms

1
New cards

What does recombinant DNA technology involve?

the transfer of fragments of DNA from one organism, or species, to another

2
New cards

What is recombinant DNA?

the DNA of two different organisms that have been combines

3
New cards

The resulting organism from recombinant DNA can be referred to as what?

transgenic

or

GMO - genetically modified organism

4
New cards

Since the genetic code is universal what does this mean for the transgenic organism?

the transcription and translation mechanisms are the same

the transferred DNA can be translated within cells of the recipient

5
New cards

What are the 3 ways fragments of DNA can be produced?

mRNA to cDNA using reverse transcriptase

using restriction enzymes to cut a fragmnet

creating the gene in a gene machine

6
New cards

What are the 5 key stages of producing transgenic organisms?

isolation of DNA fragments

insertion of DNA fragments into a vector

transformation - transfer of DNA into suitable host

Identification of host cells that have successfully taken yp the gene

growth/cloning

7
New cards

How is mRNA converted to cRNA?

mRNA isolated (cells are centrifuged)

free DNA nucleotides are added

reverse transcriptase joins DNA nucleotides to mRNA which acts as a template

enzyme (ribonuclease H) hydrolyses mRNA to get cRNA

8
New cards

is cRNA double or single-stranded?

single-stranded

9
New cards

What happens once we have cDNA?

DNA polymerase catalyses the formation of a complementary strand of DNA

produced a double strand (using free DNA nucleotides from earlier)

a copy of the gene now exists, with no introns

10
New cards

How are the cells chosen?

cell that readily produces the protein is selected

cells have large quantities of relevant mRNA which is more easily extracted

11
New cards

What do restriction endonuclease do?

cut up a DNA strand at a specific sequence of bases called a recognition sequence

12
New cards

Recognition sites are usually what?

palindromic (read the same left to right and right to left)

13
New cards

What are the different ways restriction endonuclease can cut up DNA?

some make clean cuts straight down - blunt ends

some make staggered cuts - sticky ends

14
New cards

What are oligonucleotides?

single strands of DNA

15
New cards

how does a gene machine work?

desired sequence of nucleotides can be determine by primary structure of protein

desired nucleotide base sequence is fed into a computer

the computer designs series of overlapping oligonucleotides which can be assembled into the desired gene

automated process assembles each of the oligonucleotides

oligonucleotides are joined together to make a gene - amplified by PCR

16
New cards

What happens once the gene is made?

using sticky ends, the gene can be inserted into a bacterial plasmid to be eventually cloned

17
New cards

Why can’t restriction enzymes be used to produce DNA that E.coli could be used to produce a human gene?

human DNA contains introns

E.coli cannot remove introns

18
New cards

Why is it better to use the gene machine than reverse transcriptase to convert mRNA into cDNA?

faster to use gene machine than enzyme-controlled reactions