Bio Unit 3 Sem 1 Sac Flashcards
75% (23 cards)
Polymerase
Joins nucleotides together to create nucleic acids
Taq Polymerase
Polymerase that is highly resistant to high temp - joins nucleotides together to create nucleic acid - using the original DNA template strand
DNA ligase
Join nucleic acid fragments together by helping to create the phosphodiester bonds
Restriction enzyme/ endonuclease
Cuts DNA sequences at specific sites - sticky end - (single strand overhangs) or blunt end - (straight)) cut
Plasmid
Circular DNA strand
Recombinant plasmid
Circular DNA strand combined with ampR, target gene and GFP gene
Vector (for a gene)
Transport for gene into DNA - (plasmid)
Bacterial transformation
- Obtain plasmid and target gene.
- Cut the plasmid using restriction enzyme, producing either blunt or sticky ends, then add the target gene into the plasmid using DNA Ligase to produce recombinant plasmid.
- The recombinant plasmid is then added to a bacteria cell by heat shock (very low success rate).
- Once the recombinant plasmid is in the bacteria cell, they are then placed into an antibiotic plate to kill off the bacterias that do not have the recombinant plasmid.
- The remaining bacteria colonies are then collected and grown and the proteins made by the target genes are extracted.
Sticky ends
Staggered cut resulting in overhangs of nucleotides
Genetically modified
An organism that has had it’s DNA artificially altered in any way.
Transgenic
An organism that has had a gene from a different species inserted into it genome.
GFP gene
The fluorescence gene (under UV)
ampR gene
Antibiotic Resistent gene
Properties of the genetic code
Triplet code, degenerate, universal e.g ATG
CRISPR-Cas9
The artificial version of Cas9 + sgRNA - lab made - Endonuclease + sgRNA that guides and binds to DNA/RNA to cut a section of the gene
Cas-9
Naturally made - Endonuclease that binds to DNA/RNA to cut a section of the gene
PAM
Very short segment of nucleotides (2-6) on the DNA that provides a binding site for Cas9.
Without the PAM Cas9 will not cut the DNA
Ethical approaches
Consequences-based - Aims to maximise positive outcomes and minimise negative outcomes.
Duty / Rule-based - Aims to follow set rules and responsibilities, with less regard for the consequences that may result.
Virtues-based - Aims to emphasise the moral nature of the individual - being morally good.
Ethical concepts
Integrity - the commitment to knowledge.
Justice - the commitment to fairness.
Beneficence - the commitment to maximising benefits.
Non-maleficence - the commitment to minimising harm.
Respect - the commitment to consideration.
Control Group
Group that does not receive the treatment or intervention being studied
The role of rough endoplasmic reticulum, Golgi apparatus and associated vesicles in the export of proteins from a cell via the protein secretory pathway
Rough endoplasmic reticulum - protein synthesis
Golgi apparatus - process and package protein
Vesicle - transport of proteins to outside the cell - (export)
STR: Short Tandem Repeats
Short non-coding DNA segments that vary in length between individuals making them very useful for DNA fingerprinting.
Steps of PCR
- Denaturing - DNA is heated to approximately 95°C to break the hydrogen bonds and form single-stranded DNA
- Annealing - The sample is cooled to 55°C to allow primers to bind to complementary sequences of the single-stranded DNA.
- Extention - DNA is heated to 72°C which allows Taq polymerase to work optimally. It binds to the primer, which acts as a starting point and begins synthesizing a new DNA strand.
