8.4 Gene Technology

Question 1

Where does the Vivo method to amplify DNA fragments occur?

Answer 1

-carried out inside living bodies

Question 2

What is the first step of the vivo method ?

Answer 2

-promote and terminator regions are added to DNA fragment

Question 3

what are promotor regions?

Answer 3

-DNA sequence that tells RNA polymerase when to start transcription to produce mRNA
- can only be activated by specific cell types allows it to control which body cells the proteins produced in

Question 4

what are terminator regions?

Answer 4

-tell RNA polymerase when to stop

Question 5

what is step 2 of vivo method?

Answer 5

-restriction endonuclease and ligases used to insert DNA fragments into vectors

Question 6

what is a vector

Answer 6

-Carries DNA into host cell
-plasmids or viruses/bacteriophage

Question 7

what happens with restriction endonuclease ?

Answer 7

-vector DNA and DNA fragment cut using same restriction enzyme
-vector DNA and DNA fragment have complimentary sticky ends
-(exposed bases that have complimentary base pairs)

Question 8

what’s the role of DNA ligases?

Answer 8

-joins DNA fragment to vector DNA
-forms phosphodiester bond between adjacent nucleotides on sticky ends
-condensation reaction

Question 9

what is step 3 of the vivo method?

Answer 9

-host cells transformed using vectors

Question 10

what happens if the host cell is transformed using plasmid vector?

Answer 10

-host cells persuaded to take in plasmid vector
-place in ice-cold CaCl2 solution to make cell wall permeable
-then plasmids added
-mixture is heat shocked

Question 11

How is the host cell transformed using a vector if it’s a virus/bacteriophage?

Answer 11

-infects host bacterium by injecting its DNA
-phage DNA w target gene integrates into bacterial DNA

Question 12

what is step 4 of the vivo method?

Answer 12

-marker genes used to detect genetically modified cells/organisms

Question 13

how do marker genes work?

Answer 13

-inserted into vectors at same time as target gene
-added to identify which cells have the desired gene
- as not all cells/organisms will take up the vector and be transformed

Question 14

what are 2 examples of gene markers and how do they show the cell is transformed?

Answer 14

-gene marker code for protein that give:
-resistance to antibiotic (cells surviving are transformed)
-fluorescence (glows under UV)