Manipulating Genomes Flashcards
How would you form recombinant DNA
- Restriction Enzyme used to remove gene
- Restriction enzyme used to cut vector ( plasmid agrobacteriumfaceis) and DNA Ligase joins the 2 complementary sticky ends
- Marker gene indicates which cell has took up the recombinant DNA —> gene for fluorescence changes -> examine fluorescence under UV Light
- Electrofusion
How would you identify transformed host cells
Marker genes:
Gene for fluorescence changes
Examine fluorescence change under UV light
ANTIBIOTIC RESISTANCE GROW ON AGAR
What is Polymerase Chain Reaction and state the steps
Used to AMPLIFY copies of DNA repeatedly - range of different lengths
1. Heat DNA strands at high temp so hydrogen bonds are separated between the two complementary strands
2. (Annealing) -> Primers (SHORT PIECES OF FREE DNA NUCLEOTIDES WHICH ARE COMPLEMENTARY TO BASES AT START OF FRAGMENT) attach to specific starting points on each of the separated DNA stands by forming Hydrogen Bonds
3. EXTENSION -> DNA polymerase adds free nucleotides to ends of primers extending DNA strands to form a complete copy
Why is the n. of DNA sequences after PCR not always accurate
-Radioactive labels
- Fluorescent labels
- UV Light
What are differences between somatic and germ line therapy
Somatic - CanNOT be passed down to offspring
Germ Line - Can be passed down to offspring
Somatic - Gene introduced to body
Germ line - Gene introduced to sperm
Somatic - Only some get functional gene
Germ Line - All cells get functional gene
What is somatic and germline therapy
Somatic –> Replaces mutant alleles with healthy alleles in affected somatic cells to treat diseases. Modifications not inherited
Germline –> Involves inserting a healthy allele into germ cells or embryos to prevent genetic diseases from birth. Modifications able to be passed down to future generations
What is pharming
Process of pharmaceutical drugs produced from genetically modified organisms (Ex - goats can be genetically engineered to produce antithrombonin)
What are adv. and disadv. of pharming
Adv . - Enables mass production
- Makes new drugs accessible
Disadv. - Raises animal welfare concerns
-Can lead to animals being viewed as solely as commodities
What are adv. and disadv. of GM pathogens
Adv. - Offers potential treatment for disease previously deemed incurable
-Useful in creating vaccines
Disadv. - Carries risk of accidental infection + disease outbreak
Can be misused in biological warfare
What is electrophoresis
Putting DNA pieces in size order -> to read base sequence. Smaller molecules travel further and faster and vice versa. Negatively charged DNA moves towards the anode
what is dna sequencing
Working out sequences of bases in DNA
Why is only selected sections of non-coding DNA used when profiling
- Human genome is similar in most people
- Coding regions would not provide a unique profile
- Non coding regions contain variable number of repeating sequences
Why is the binding of negative charge to proteins necessary in electrophoresis
- Different proteins have different charges
- So all proteins have a negative charge
- So all proteins can travel in the same direction
what can mRNA do after being isolated with a restriction enzyme
- makes cDNA (single stranded DNA) using reverse transcriptase
- makes double stranded DNA using DNA Polymerase
What is adv. of bioinformatics
- Facilitated access to large amounts of data
- Facilities access to data on DNA and proteins
- Format is universal hence can identify source of outbreak
