In Vivo Gene Cloning - the use of vectors 1 Flashcards
What is used to locate a specific DNA fragment containing the required gene?
A DNA probe
What are the two main methods of gene cloning?
In vivo, which is transferring the fragment of the required gene to a vector and in vitro using the polymerase chain reaction
Why is it necessary to clone a DNA fragment once it has been identified?
To produce a sufficient quantity for medical/commercial use
What are the sequences of DNA cut by restriction endonucleases called?
Recognition sites
What happens if a restriction endonucleases cuts DNA in a staggered fashion?
The cut ends are left with single - stranded sections of DNA of a few nucleotide bases long
Why are the single - stranded sections of DNA left after cutting complementary to each other?
This is because they were previously paired together in the double strand before being cut
What happens when the same restriction endonucleases is used to cut DNA?
All fragments produced will have complementary sticky ends, allowing them to be joined together
What enzyme is used to join sticky ends together?
DNA ligase
How does DNA ligase join DNA fragments?
It binds the phosphate - sugar backbone of the two DNA sections, uniting them as one
What is the function of a promoter region in DNA transcription?
It is the binding site for RNA polymerase and transcription factors, allowing transcription to begin
What is the function of a promoter region in DNA transcription?
Why must a promoter be added to a DNA fragment?
It is the binding site for RNA polymerase and transcription factors, allowing transcription to begin
Without a promoter, RNA polymerase cannot bind and gene will not be transcribed into mRNA
What is the function of a terminator region in DNA transcription?
Why must a terminator be added to a DNA fragment?
It signals RNA polymerase to detach from the DNA, ending transcription
To ensure transcription stops at the correct point and does not continue beyond the intended gene
What enzymes synthesis mRNA during transcription?
RNA polymerase
How do transcription factors assist in gene transcription?
They help RNA polymerase bind to the promoter region initiating transcription
