Biotechnology final study guide

Archaea, Fungi, Protozoa, viruses, bacteria, algae

Streak plate method, spread plate method

Start with a swab and spread left to right. Stop. Continue from where you stopped

pour mixture on plate, spread, let colonies grow

- Cell wall

-made up of peptidoglycan

circular/oval

-can carry plasmid

- flagellum - pilli

- cell wall - chromosome

- cytoplasm - ribosome

cells that are able to take up DNA, when exposed to calcium chloride (CaCl2), and heat shocked

To mix competent bacterial cells and DNA in solution with CaCl2

To put the mixture in ice so the DNA sticks to the bacterial wall

Heat mixture to 37-42 degrees C and let DNA enter the now competent bacteria cell wall through the open pores

Plasmid DNA is replicated by bacteria cells, transcribed into mRNA, and translated into protein & cloned human proteins

is a mixture of bacteria and recombinant Plasmid DNA with a gene for antibiotic resistance

Transfer mixture an electroporation cuvette

You apply an electrical shock so the bacteria opens up and lets the DNA enter it

Move the cells to an antibiotic plate and select the cells that took up the recombinant DNA

N-Terminal has (NH2-) at the end of the peptide with an amine group

C-terminal has (-CooH) at the end of the peptide with carboxyl group.

N--------C

When a target protein is fused to a bacterial protein

It treats type 1 & 2 diabetes

To lower glucose in the blood

You produce chain A and chain B separately in a plasmid both containing

- a promoter

-B - galactosidase gene

- antibiotic resistance gene

The B-galactosidase becomes fused with the chains

Transform Bacterial cell (E. coli) so the fused A/B chain with the B-gala can enter the cell

Purify it so you're left with the A/B chain fused with the B-gala

Chemically remove the Insulin Chain from the B-gala and connect it with the opposite chain. The insulin then becomes active once combined

This creates a

NH2 ----- COOH (A protein chain)

NH2 ------COOH (B protein chain)

Lux Genes

it releases light when it is being converted from luciferin to oxyluciferin (produces light in the presence of oxygen)

When Lux Gene ecodes Luciferase that uses O2 and (ATP) which converts lluciferin to oxyluciferin which releases light.

Lux gene --- Luciferin + ATD + O2 ----- Light ---- oxyluciferin + AMP + CO2

Is used in PCR

Thermostat able DNA Polymerase

Can wistand high temp. used in cycling

From thermus aquatics

THERMOPHILIC BACTERIUM

substances produced by one microorgansim that inhibit the growth of bacteria

-Different mechanism kills bacteria directly or inhibit bacterial population

Isolated from bacteria then inhibit the growth of it

Ex:

Penecillin

Tetracycline

They prevent it from creating a cell wall

Then inhibit the cell wall

Damage plasma membrane

Prevent enzymes that require cell metabolism.

Glucose uses

NAD+ (Low energy) which reduces to NADH (High energy)

2 ADP (Low energy) to make 2 ATP (High energy)

goes through glycolysis

Creating Pyruvate which uses

NADH to make NAD+

goes through lactic fermentation

Makes Lactic Acid

Glucose uses

NAD+ which reduces to NADH

2 ADP to make 2 ATP

goes through Glycolysis

Creating Pyruvate which uses

NADH to make NAD+

goes through alcohol fermentation

Creates ethanol + CO2

Developed: 1796 ; Edward Jenner

An 8 yr old was vaccinated with live cowpox (from skin ulcers)

Was then given small pox 9 days later

Cowpox = Protection from small pox

Since cowpox and smallpox are closely related viruses, his immune system developed enough protection to fight smallpox.

So effective it eradicated the disease in 1980

That the Measles, Mumps, & Rubella (MMR) vaccine caused Autism

Which is false

Gardasil and Hepatitis B

Liver cancer

Human Papillomavirus (HPV)

Cervical, vaginal, vulvar, throat, anal, & penile cancer

Antibodies that have diminished death rates

The (DSMB) had to monitor the safety of the antibody

FDA had to review and vote (FDA Approved)

Bind to the antigen and target invader for destruction (KILL)

non-self molecules present on the surface of bacteria, viruses, fungi, or any other source that elicit antibody production (anything that causes immune system response)

Recognizing specific antigens

Targeting that antigen

Getting rid of pathogen

phagocytes

lymphocytes

B cells

T cells

Cytotoxic (CD8+)

Helper (CD4+)

Engulf the antigen

Are activated by antigens binding to cell

consist of B and T cells

Kill/ destroy pathogen/antigen

Cytotoxic (CD8+) - To bind to the infected cell and attack/kill it

Helper T (CD4+) - Releases cytokines to activate other cells

Make antibodies to attack outside the cell (must be activated by helper T)

Top : Variable domain (antibody binding site)

Middle: Light chain

Bottom: Heavy chain (Constant domain)

Occurs the first time an individual is exposed to a specific antigen. Production of antibodies in this case takes a few days. B cells are activated to form Plasma cells (that produce antibodies) and Memory cells (that present the antibodies on their surface).

Occurs as the result of a subsequent exposure to a specific antigen. Response is greatly enhanced over that of the primary response and utilizes Memory cells produced in the primary response. This high level of resistance is known as "active immunity".

A secondary response through previous infection

by getting exposed to a virus or vaccine

Body making it

Injection

Becoming immune from someone or something else

Mother - baby (milk)

Medicine

Attenuated Vaccines

Inactivated Vaccines

Subunit Vaccines

Genetic Vaccines

dead microbes injected into a person so the body becomes immune to it

Virus cannot replicate

(Flu Vaccine)

Using live viruses that are modified and cannot replicate

(Chickenpox)

Uses the protein on the pathogen surface to make a vaccine by using recombinant DNA Tech

(Hepatitis B)

Add mRNA or DNA for viral proteins. (Use host system to express protein from genetic material.

Not exposed to virus (mRNA cells produce viral protein)

Mutates Rapidly - Has many variants

Has a RNA genome

Uses reverse transcriptome to make DNA copy from the RNA after infecting host cell

Insert the DNA into the genome of host cell

Target molecules that are required to replicate the virus

when used in a mixture of different antiviral drugs

Target the viral enzyme : Reverse Transcription, integrase, & HIV - 1 protease.

Timothy Ray Brown

was diagnosed with HIV (acute myeloid leukemia)

Treated in 2007 with BMT (Bone Marrow Transplant that produces T cells that could not be infected by the virus)

Died sep 29, 2020 from leukemia

Mutates rapidly

Has RNA genome,

no DNA (Intermediate)

H - Hemagglutinin (HA)

attacks cells

antibodies target it

N - Neuraminidase (NA)

allows replicated virus to exit cells

tamiflu targets it

Spike protein

Moderna

Pfizer

Johnson & Johnson (J & J)

Astra Zeneca

Protein Based subunit

Use mRNA

90% effective against disease

Uses virus to deliver dna to cell 72 - 90% efficacy

Novauax 89% efficacy

In eggs

- Inactivated (Flu shot)

In cultured cells

- virus replicated in mammalian cells instead of eggs (grow for a while)

HA protein grown in insect cells and isolated

- 100 % egg free

mRNA for the spike protein is produced in dish/test tube

mRNA is packaged in lipid/nanoparticle so it is able to be delivered

Particles used in vaccine

Uses randomly generated short primers to produced distinct sets of PCR for a given organism

network of labs use DNA fingerprinting to rapidly identify strains of microbes important to public health

Electrophoresis: a version of agarose gel electrophoresis that has been modified for use with large pieces of DNA

Mass of plant that can form any type of cell in the plant, and can divide and form a whole plant

A plant cell treated to remove the cell wall

auxin = root growth

cytokinin = shoot growth

organelle where photosynthesis takes place (Location of chlorophyll)

1) Dissect leaf and soak it with cellulase, sugar, salt

2) Let it sit in liquid the protoplast will float up while cell wall sinks

3) Isolate protoplast in an agar with nutrients, culture it and induce fusion from different plants

4) Transfer callus to medium so the plants grow

Transgene is inserted into the DNA of the chloroplast instead of nuclear DNA.

You have an engineered Ti plasmid from agrobacterium that can transfer plasmid into plant cell

That plasmid induced plant cells to make food molecules

Higher expression levels

Limits cross contamination

Can add several genes in tandem

1) double stranded RNA is cut into small fragments by dicer and becomes (siRNA)

2) RISC then gets siRNA (single strand) and matches it with mRNA

3) Slicing it then done by RISC to the mRNA

4) RNA is then destroyed as well to prevent from mRNA from becoming protein

Come back to it

1) Scientist got an egg cell from a doner and removed the nucleus from it

2) They got a nuclear/nucleus doner from a different sheep

The egg cell and the nucleus were combined through electric shock (the egg cell opens so single cell enters)

The egg cell now has a nucleus

The cells then divide and become blastocyst

The blastocyst is then inserted in the uterus of a sheep surrogate

Doesn't produce an exact duplicate

Doner cell must come from a living organism

Mitochondrial DNA is different

Different patter of X inactivation (if female)

Oxidation = loss of an electron

Reduction = gain of an electron

An electron doner goes through oxidation and loses an electron becoming an oxidized doner (+)

An electron acceptor goes through reduction gains an electron and becomes a reduced acceptor (-)

Presence of O2

Benzene transfers electrons to O2 and becomes CO2 + H2O + biomass

O2 is not present

Toluene transfers electrons to NO3- and becomes CO2 + N2 + H2O + biomass

Toluene is oxidized to CO2

Carbon Dioxide (CO2)

Methane (CH4)

Nitrous oxide (N2O)

Ozone (O3)

Extra: Fluorinated gases

Facilitates the binding of transcription factors and RNA pol for gene transcription

Specific region of a gene where RNA polymerase can bind and begin transcription

A protein that suppresses the transcription of a gene

Bind to repressors and turn off transcription

A system in which a repressor is bound to an operator, and an inducer is required to activate it by preventing the repressor from binding to the operator

Characteristics:

- Lactose = energy source for Prokaryotes

- Lac Repressor binds to operator and prevents RNA pol from binding

- Lactose = Inducer (binds to repressor and induces transcription)

- Need CRP to bind to cAMP (cyclic AMP) to get RNA pol to bind, and glucose regulates CRP

- High glucose= no CRP and cAMP to recruit RNA pol

Used to block translation or chew up RNA post-transcription, which aids in gene editing

An enzyme that cuts DNA by attaching to a guide RNA, base-pairing with DNA, and cleaving

Uses:- Can be used to alter, add, or cut genes post-transcription

DNA molecule made in a lab that is derived from at least two genetic sources

three/four

small trials and the safety of it