Topic 5: Recombinant DNA Technology + Topic 6: Reproductive Biotechnology and Immunology
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80 Terms
Q: What is a vector?
A DNA molecule used to transfer DNA from one organism to another.
Q: What are plasmids
Small circular double-stranded extrachromosomal DNA used as vectors.
Q: What is the function of ori?
Allows independent replication of a plasmid.
Q: What is a multiple cloning site (MCS)?
Region with restriction enzyme sites for DNA insertion.
Q. What are selectable markers?
Genes used to identify transformed cells (e.g. ampR, tetR).
Q: What is a reporter gene?
Gene that produces a detectable signal to indicate gene expression.
Q: What does lacZ do?
Used in blue-white screening.
Q: What colour indicates successful insertion in lacZ screening?
White.
Q: What are bacteriophages used for?
As vectors to carry DNA into bacteria.
Q: What are COS sites?
Sticky ends that allow circularisation of phage DNA.
Q: What are plaques?
Zones of dead bacteria caused by phage infection.
Q: What is a cosmid?
Hybrid vector (plasmid + phage).
Q: What is a BAC?
Stable vector carrying large DNA (100–300 kb).
Q: What is a YAC?
Vector carrying very large DNA (200–2000 kb).
Q: What are expression vectors used for?
Producing proteins.
Q: Why can’t bacteria produce complex proteins properly?
They lack post-translational modifications.
Q: What are viral vectors?
Viruses used to deliver DNA into cells.
Q: How do retroviruses work?
RNA → DNA → integrates into host genome.
Q: What is the result of retroviral integration?
Permanent gene expression.
Q: What is transformation?
Uptake of DNA by bacteria.
Q: What is electroporation?
Electrical pulses create pores for DNA entry.
Q: What is a gene gun?
DNA-coated particles shot into cells.
Q: What is transfection?
DNA delivery into eukaryotic cells.
Q: What is microinjection?
Direct injection of DNA into a nucleus.
Q: What is blotting?
Technique to detect DNA, RNA, or proteins.
Q: What does Southern blot detect?
DNA.
Q: What does Northern blot detect?
RNA.
Q: What does Western blot detect?
Proteins
Q: What is PCR?
Technique to amplify DNA.
Q: What are the steps of PCR?
Denaturation, annealing, extension.
Q: What is qPCR?
Real-time DNA quantification using fluorescence.
Q: What is Sanger sequencing?
Method to determine DNA sequence using chain termination.
Q: What are ddNTPs?
Nucleotides that stop DNA synthesis.
Q: What are microarrays used for?
Measuring gene expression of many genes at once.
Q: What is artificial insemination (AI)?
Artificial introduction of semen into the female reproductive tract.
Q: When does sperm damage occur?
During freeze/thaw.
Q: What are the two cloning methods?
Embryo splitting and nuclear transfer.
Q: What is embryo splitting?
Artificial twinning producing identical individuals.
Q: What is nuclear transfer?
Replacing egg nucleus with donor nucleus.
Q: Why is Dolly important?
Proved adult cells can be reprogrammed.
Q: What stage were donor cells placed in for Dolly?
A: G₀ stage.
Q: What is MOET?
Multiple Ovulation and Embryo Transfer.
Q: What is conservation biology focused on?
Preserving genetic diversity.
Q: What is innate immunity?
Non-specific, no memory.
Q: What is adaptive immunity?
Specific with memory.
Q: What are phagocytic cells?
Cells that engulf pathogens.
Q: What is opsonisation?
Tagging pathogens for phagocytosis.
Q: What is an antigen?
Molecule that triggers immune response.
Q: What do B cells do?
Produce antibodies.
Q: What do T cells do?
Kill infected cells or activate immune responses.
Q: What is Major Histocompatibility Complex MHC?
Proteins that present antigens to T cells.
Q: What are antibodies?
Proteins that bind antigens.
Q: What is the complement system?
Protein cascade that destroys pathogens.
Q: What does histamine do?
Increases blood flow and permeability.
Q: What are vaccines?
Introduce antigen to create memory cells.
Q: Difference between monoclonal and polyclonal antibodies?
Monoclonal = one epitope; polyclonal = multiple epitopes.
Q: What is ELISA?
Test that detects antigen or antibody using colour change.
Q: What is Western blotting used for?
Detecting proteins.
Q: What is PCR used for (applications)?
Disease detection, genetic mutation analysis, forensic DNA profiling, cloning, and sequencing preparation.
Q: What is qPCR used for?
Measuring gene expression levels by quantifying DNA in real time.
Q: What is Sanger sequencing used for?
Determining the exact DNA sequence and identifying mutations.
Q: What are microarrays used for?
Measuring expression of many genes at once and comparing gene activity (e.g. healthy vs diseased cells).
Q: What is ELISA used for?
Detecting and quantifying antigens or antibodies (e.g. disease testing).
Q: What are reporter genes used for?
Studying gene expression and promoter activity by producing a detectable signal.
Q: What are the two main types of immune response?
Innate (natural) and adaptive (acquired).
Q: What are the two types of adaptive immunity?
Humoral immunity and cell-mediated immunity.
Q: What is humoral immunity?
B cell response that produces antibodies to target pathogens.
Q: What is cell-mediated immunity?
T cell response that destroys infected or abnormal cells.
Q: What are cytokines?
Signalling molecules that coordinate and regulate immune responses.
Q: What do cytokines do overall?
Control communication between immune cells and direct immune activity.
Q: What are interferons?
Cytokines released by infected cells that warn neighbouring cells and trigger antiviral responses.
Q: What are interleukins?
Cytokines that allow communication between immune cells and stimulate immune responses.
Q: Difference between cytokines, interferons, and interleukins?
Cytokines = general signalling molecules
Interferons = antiviral signals
Interleukins = communication between immune cells
Q: What is the purpose of vaccines?
To stimulate an immune response and create memory cells for faster protection.
Q: What are the main types of vaccines?
Live attenuated, inactivated, subunit, and recombinant.
Q: What are live attenuated vaccines?
Weakened forms of the pathogen that still trigger immunity.
Q: What are inactivated vaccines?
Killed pathogens that cannot replicate.
Q: What are subunit vaccines?
Only parts of the pathogen (antigens) are used.
Q: What are recombinant vaccines?
Vaccines made using genetic engineering to produce antigens.
Q: Why are vaccines effective long-term?
They create memory B and T cells for a faster secondary response.