Biotechnology: Principles and Processes

$94^{\circ}C$

$40-65^{\circ}C$

$72^{\circ}C$

$100^{\circ}C$

Polymerase

Plasmid

Primer

Pathogen

Upstream processing

Downstream processing

Transformation

PCR

Maintain a constant pH

Ensure oxygen availability throughout the reactor

Add nutrients to the medium

Prevent the growth of contaminants

Terminating the process of translation.

Initiating replication and controlling the copy number of linked DNA.

Binding the RNA polymerase to the promoter region.

Protecting the DNA from being cut by restriction enzymes.

They created a new species of bacteria called $E$. $colimonella$.

They demonstrated that DNA from different sources could be joined and function in a host.

They proved that antibiotics cannot kill bacteria containing plasmids.

They showed that DNA can only replicate in its original organism.

Tissue culture of non-modified cells.

Development of recombinant DNA molecules.

Observation of phenotypic changes in the wild.

Application of chemical fertilizers to soil.

A bacteriophage protein coat

A plasmid from Salmonella typhimurium

A modified mRNA molecule

A ribosome from E. coli

They are isoschizomers.

They produce identical overhanging sequences despite having different recognition sites.

The enzymes were isolated from the same species of bacteria.

Both enzymes produce blunt ends.

They neutralize the negative charges on both the DNA and the bacterial cell wall, facilitating entry.

They act as a cofactor for the DNA polymerase inside the host cell.

They cause the DNA to precipitate onto the surface of the cell.

They break the phosphodiester bonds of the plasmid to allow it to enter.

Capacity

Competence

Virulence

Efficiency

DNA Ligase

Restriction Endonuclease

DNA Polymerase

Primase

$5'-CTTAAG-3'$

$5'-GAATTC-3'$

$5'-TTAACG-3'$

$5'-GAAUUC-3'$

Color of the fragments

Charge only

Size or length of the fragments

Temperature of the fragments

The two strands of the template DNA are separated by heat

Primers bind to their complementary sequences on the single-stranded DNA

Taq polymerase adds dNTPs to the $3'$ end of the growing chain

The reaction is cooled to $0^\circ C$ to stop all enzymatic activity

Cellulase

Lysozyme

Lipase

Protease

It confers resistance to the antibiotic streptomycin

It codes for proteins involved in the replication of the plasmid

It acts as a site for the insertion of foreign DNA

It is responsible for the secretion of the recombinant protein

They consist of double-stranded DNA segments.

They facilitate the action of DNA ligase by forming temporary hydrogen bonds between complementary bases.

They prevent the formation of recombinant DNA molecules.

They are only produced by Type I restriction enzymes.

To denature the DNA and make it single-stranded.

To create temporary pores in the bacterial cell wall/membrane to allow DNA entry.

To activate the antibiotic resistance genes in the plasmid.

To kill any non-recombinant bacteria in the mixture.

No enzyme is used; it is the physical removal of DNA from the gel piece.

DNA Ligase

Lysozyme

Restriction Endonuclease

Only traditional views

Only modern molecular biotechnology

Both traditional views and modern molecular biotechnology

Only plant tissue culture

Cellulase

Lysozyme

Chitinase

Amylase

It makes the vector unstable

It simplifies the cloning process by preventing fragmentation of the vector

It allows for faster replication

It helps in selecting only large genes

Join DNA strands

Synthesize DNA

Cut DNA at specific locations

Degrade the cell wall

It is a natural plasmid found in E. coli

It was the first artificial cloning vector constructed by Bolivar and Rodriguez

It lacks an origin of replication

It cannot be used to clone DNA fragments larger than 100 bp

The 5' to 3' direction on one strand being identical to the 3' to 5' direction on the same strand

The 5' to 3' direction on one strand being identical to the 5' to 3' direction on the complementary strand

The ratio of Purines to Pyrimidines

The presence of a promoter sequence

It makes the plasmid twice as stable

It allows for the selection of recombinants by insertional inactivation of one of the genes

It ensures the plasmid can replicate in two different host species

It prevents the bacterial host from dying in any antibiotic environment

The pores become so small that even small DNA fragments move very slowly or not at all.

The DNA fragments will denature into single strands due to the heat of the gel.

The gel will not solidify, preventing the formation of wells.

The DNA will bind irreversibly to the agarose and cannot be visualized.

RNase H

Lysozyme

Exonuclease III

Alkaline Phosphatase

To allow the bacteria time to express the antibiotic resistance genes before they are exposed to the antibiotic.

To wash away the calcium chloride which would otherwise inhibit the growth of the bacteria.

To ensure that the plasmid DNA has completely integrated into the bacterial chromosome.

To provide a chance for the bacteria to repair the damage caused by the high temperature of $42^{\circ}C$.

Proteins

Lipids

DNA

Carbohydrates

DNA Polymerase I

DNA Polymerase III

Taq polymerase

RNA Polymerase

Stop the growth

Keep the cells in stationary phase

Keep the cells in their physiologically most active log phase

Decrease the product yield

To amplify the gene of interest

To convert raw materials into a finished product

To separate and purify the desired protein

To insert DNA into a host cell

Centrifugation or Filtration

Ligation

Polymerization

Annealing

High levels of blood glucose

Low concentrations of a pathogen (like a virus) before symptoms appear

The presence of vitamins in a food sample

The total white blood cell count

Only the upstream gene insertion

Only the fermentation stage in the bioreactor

Both the biosynthetic stage and the downstream processing

Only the final packaging and marketing

It remains constant.

It increases, making oxygen supply easier.

It decreases, making oxygen supply more challenging.

It becomes irrelevant because of the impeller.

The reaction would still work but slower.

No amplification would occur because the primers cannot bind at $72^{\circ}C$.

The polymerase would be inactivated.

The DNA would remain single-stranded.

Purification $\rightarrow$ Cell Lysis $\rightarrow$ Filtration $\rightarrow$ Formulation

Cell Lysis $\rightarrow$ Filtration $\rightarrow$ Purification $\rightarrow$ Formulation

Filtration $\rightarrow$ Purification $\rightarrow$ Cell Lysis $\rightarrow$ Formulation

Formulation $\rightarrow$ Cell Lysis $\rightarrow$ Purification $\rightarrow$ Filtration

Bacteria

Plants

Animals

Viruses

To increase its growth rate

To remove its ability to cause tumors while keeping its DNA delivery ability

To make it produce more oxygen

To make it resistant to heat

Hydrophobic

Hydrophilic

Soluble in lipids

Too small

Low velocity

Zero velocity

High velocity

Variable velocity

Escherichia coli

Streptococcus pneumoniae

Agrobacterium tumefaciens

Salmonella typhimurium

It should be easy to grow and maintain

It should have a very slow growth rate

It should naturally reject any foreign DNA

It should be a multicellular organism only

Into the cytoplasm

Directly into the nucleus

Into the mitochondria

Into the cell wall

High salt levels would cause the DNA to precipitate out of the solution.

High salt levels increase the conductivity of the medium, which can lead to electrical arcing and cell death.

Salts inhibit the formation of pores by stabilizing the lipid bilayer.

Deionized water helps to swell the cells, making them easier to puncture.

The plasmid DNA itself.

The gold or tungsten particles coated with DNA.

The plastic disk that holds the particles.

The small bacterial cells used to deliver the DNA.

Any bacterial strain can be used as a host for any plasmid vector.

The host must be 'restriction-deficient' ($r^-$) to ensure the incoming foreign DNA is not digested.

The host must have a higher GC content in its genome than the plasmid to ensure stability.

The host cell must be in the stationary phase to maximize the uptake of DNA.

Synthesis

Degradation

Recovery

Infection

Yes, it is identical

No, it varies from product to product

Yes, it only involves tasting

No, it only involves color checks

The height of the scientist

The nature and purity required for the product

The color of the laboratory

The shape of the petri dish

In a crude form

In a refined and finished form

Mixed with bacteria

In a gaseous state

To remove the biomass from the culture broth

To increase the mutation rate

To insert the gene into the plasmid

To provide nutrients to the bacteria

Counting the number of cells in the bioreactor

Documented evidence that a process consistently produces a result meeting predetermined specifications

Checking if the computer is turned on

Measuring the weight of the raw materials

DNA polymerase

Salts like ammonium sulfate

Restriction enzymes

Agarose gel

Less than 1000 CFU per mL

Zero (Sterile)

Exactly 10 CFU per mL

Bioburden does not matter if the product is effective.

General Metabolic Pathway

Good Manufacturing Practice

Genetic Modification Protocol

Gross Molecular Purity

Phenol or m-Cresol

Ethidium Bromide

Hydrochloric Acid

Sodium Hydroxide