Gene Technology Flashcards
What is recombinant DNA technology?
Combining of different organism DNA which could enable scientists to manipulate and alter genes to improve industrial processes and medical processes
What 3 methods of obtaining DNA fragments are there?
Reverse Transcriptase
Restriction Endonucleases
Gene machine
How does the enzyme reverse transcriptase work?
Reverse transcriptase joins DNA nucleotides with complementary bases to mRNA sequence.
Single stranded DNA is made (cDNA)
To make DNA fragment double stranded enzyme DNA polymerase is used
Is cDNA intron free and why?
Yes because it is based on the mRNA template, mRNA has introns removed.
What are restriction endonucleases?
Enzymes that cut up DNA
What effect of enzymes cutting to create staggered ends?
Staggered ends created known as sticky ends because they have the ability to join to DNA with complementary base pairs due to their exposed DNA bases
In the Gene machine once the mRNA and DNA sequences have been identified what is the name given to small sections of overlapping strands of nucleotides?
Oligonucleotides
What can be oligonucleotides be used for?
To create DNA for the entire gene
What can be used to amplify the quantity and to make a double strand from the oligonucleotide?
PCR
Name 3 advantages of the gene machine
Quick
Accurate
Makes intron free DNA so be transcribed in prokaryotic cells
Why are gene machines more useful than other techniques for producing DNA fragments?
A DNA template isn’t necessary
What sections of DNA do restriction endonucleases attach to?
Recognition sequences
What is in Vivo cloning?
Where copies of the DNA fragment are made inside a living organism
What is a vector?
Something that is used to transfer DNA into a cell
What are 2 examples of Vectors?
Plasmids
Bacteriophages
What is the role of DNA ligase in Vivo cloning?
DNA ligase joins the sticky ends of the DNA fragment to the sticky ends of the vector DNA- process known as ligation
What does vector DNA + DNA fragment produce
Recombinant DNA
What is a vector with the recombinant DNA used for?
Used to transfer the gene into cells (host cells)
If a plasmid vector is used, how does it enter the host cell?
Host cells have persuaded to take in the plasmid vector and its DNA
If a bacteriophage vector is used, how does it enter the host cell?
The bacteriophage will infect the host bacterium by injecting its DNA into it. The phage DNA (with the target gene in it) then integrates into the bacterial DNA
What is the name given to host cells that take up the vectors containing the gene of interest?
They are said to be transformed
What can be used to identify transformed cells?
Marker genes can be used to identify transformed cells
How are marker genes used?
Inserted into vectors at the same time as gene to be cloned
Transformed cells will produce colonies where all the cells contain the cloned gene and the marker gene
Marker gene can code for specific antibiotic resistance so only transformed cells with the marker gene will survive and clone or it can code for fluorescence when the Algar plate is placed under a UV light only transformed cells will fluoresce
Identified transformed cells are allowed to grow more, producing lots and lots of copies of the cloned gene
What are promotor regions?
DNA sequences that tell the enzyme RNA polymerase when to start producing mRNA.
